JP3418762B2 - Amide derivatives of antibiotic A40926 - Google Patents

Abstract

The present invention is directed to novel antibiotic A 40926 derivatives characterized by having a carboxy, (C1-C4)alkoxycarbonyl, aminocarbonyl, (C1-C4)alkylaminocarbonyl, di(C1-C4)alkylaminocarbonyl or hydroxymethyl substituent on the N-acylaminoglucuronyl moiety and a hydroxy or a polyamine substituent in position 63 of the molecule. The compounds of the invention show high in vitro activity against glycopeptide resistant Enterococci and Staplylococci.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to antibiotic A40926 derivatives of formula (I) and pharmaceutically acceptable addition salts thereof.

In the above formula, R₁Is a protective group for hydrogen or an amino functional group.
Representation; R₂Is (C₉-C₁₂) Represents alkyl; M is hydrogen, α
-D-mannopyranosyl or 6-O-acetyl-α
Represents -D-mannopyranosyl; Y is carboxy, (C₁
-C_Four) Alkoxycarbonyl, aminocarbonyl, (C₁
-C_Four) Alkylaminocarbonyl, di (C₁-C_Four) Archi
Luaminocarbonyl [These alkyl moieties are hydro
Xy, amino, (C₁-C_Four) Alkylamino and di (C₁
-C_Four) Having one substituent selected from alkylamino
May be. ] Or hydroxymethyl; X
Is a hydroxy or formula -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃Is hydrogen or (C₁-C_Four) Represent alkyl
Shi; alk₁, Alk₂And alk₃Are each independently a carbon atom 2
Represents ~ 10 linear or branched alkylenes; p and q
Is independently an integer representing 0 or 1; R_FourAnd R
_FiveAre each independently hydrogen, (C₁-C_Four) Represents alkyl
Or R₃And R_FourTogether is the article where p is 1
Linked to two nitrogen atoms (C₂-C_Four) Arche
Or R_FourAnd R_FiveTogether p
Linked to two nitrogen atoms under the condition that both q and q are 1
(C₂-C_Four) Represents an alkylene moiety; W is hydrogen, (C₁
-C_Four) Alkyl, amino, (C₁-C_Four) Alkylamino,
The (C₁-C_Four) Alkylamino, 1 or 2 amino
− (C₂-C_Four) Alkyl moieties or 1 or 2 (C₁
-C_Four) Alkylamino- (C₂-C_Four) Alkyl moieties or
1 or 2 di (C₁-C_Four) Alkylamino- (C₂−
C_Four) Represents amino substituted by an alkyl moiety,
Alternatively, if p and q are both zero, then W is
Partial-NR₃−alk₁-Together with Piperazino
May represent 4-methylpiperazino] In the case where X represents hydroxy,
In, Y represents hydroxymethyl, Z is hydrogen or
Is based [In the formula, A Represents an anion of an inorganic or organic acid
Or carboxylic acid functionality left on the antibiotic
If present in the part, it also includes the carboxylic acid
May represent an internal anion derived from a functional group] Represents

The numbers in parentheses in the above formula (I) and the following formulas are conventional numbers of relative carbon atoms in the molecular structure of antibiotic A40926 and its derivatives.

Antibiotic A40926 is a glycopeptide antibiotic complex obtained by culturing Actinomadura, designated Actinomadura sp. ATCC39727, in a medium containing assimilable carbon, nitrogen sources, and inorganic salts. (See European Patent No. 177882). According to the method described in the cited patent, recovery of the present antibiotic complex whose main factors are named Factor A, Factor B, Factor B ₀ , Factor B ₁ , Factor PA, and Factor PB is Filtering or pre-purifying the fermentation broth followed by affinity chromatography on immobilized D-alanyl-D-alanine.

The as-identified factors of this A40926 can be represented by the following formula (II).

In the formula, R ′ ₁ is hydrogen, X ′ is hydroxy, and Y ′
Is carboxy, R ′ ₂ represents a (C ₉ -C ₁₂ ) alkyl group, and M ′ represents an α-D-mannopyranosyl or 6-O-acetyl-α-D-mannopyranosyl group.

More particularly, the antibiotic A40926 Factor A is one compound of formula (II) above, in which R ′ ₁ is hydrogen, X ′ is hydroxy, Y ′ is carboxy and R ′ is ₂ represents n-dodecyl, and M'is α-D
-Represents mannopyranosyl. According to the most recent research
Antibiotic A40926 described in European Patent 177882 above
A substance identified as Factor B consists of two components that are substantially in close relationship. Antibiotic A40926 factor
B ₀ is in fact the main constituent of factor B and corresponds to the compound of formula (II) above, in which R ′ ₁ is hydrogen, X ′ is hydroxy and Y ′ is carboxy. , R '
₂ represents 9-methyldecyl and M ′ represents α-D-mannopyranosyl.

The smaller component of Factor B is termed Factor B ₁ and differs from Factor B ₀ only in that R ′ ₂ represents n-undecyl (E. Riva et al, Chromatographi.
a, Vol. 24, 295, 1987).

Antibiotics A40926 Factors PA and PB differ from the corresponding Factors A and B in that the mannose units are replaced by 6-O-acetyl-α-D-mannopyranose units.

Antibiotics A40926 Factors PA and PB, under at least some fermentation conditions, are the major antibiotic products of the microorganism producing A40926.

Antibiotics A40926 Factors A and B are
It is primarily a conversion product of the antibiotics A40926 factors PA and PB, and is often already present in the fermentation broth.

All sugar moieties are linked to the nucleus of antibiotic A40926 through O-glycosidic bonds.

Under basic conditions such as removing the acetyl group of the mannose unit without substituting the acyl group for the aminoglucuronyl unit, the antibiotic A40926 Factor PA can be converted to the antibiotic A40926 Factor A, and A4
It was discovered that Factor PB can be converted to the antibiotic A40926 Factor B.

Subsequently, the fermentation broth or the extract containing the antibiotic A40926 or its concentrate is kept under basic conditions for a certain period of time (for example,
When left in an aqueous solution of a nucleophilic base at pH 9 or higher (overnight), an antibiotic A40926 complex containing antibiotics A40926 Factor A and Factor B is obtained.

The antibiotic A40926 Factor B can be obtained from the antibiotic A40926 complex by chromatographic separation using the method described in EP 177882. Under the conditions described in the above European patent, about
Pure Factor B ₀ , described as 90%, can be obtained by further purification of Factor B, eg by repeated, reverse phase chromatography.

More recent research (L.Zerilli et al., Rapid Communi
cations in Mass Spectrometry, Vol.6,109,1992), in the antibiotic complex A40926, and also in acronyms A ₁ , RS-1, RS-2 and RS-, respectively.
It was found that there were several trace factors identified as 3. These minor factors were individually isolated by HPLC and their structures were determined by applying gas chromatography / mass spectrometry of the methanolysis of the A-40926 complex.

All of the above-mentioned trace factors have a structure corresponding to the basic structure of factors A, B ₀ and B ₁ apart from the fatty acid residue linked to the aminoglucuronic acid moiety. More preferably, when referring to Formula _{(II), R '1,} X' and Y 'have the meanings same as the above, whereas R' ₂ is the factor A ₁ 8- methylnonyl, factor RS-1 Represents n-nonyl and Factor RS-3 represents n-dodecyl.

In the antibiotic A40926 complex preparations generally obtained by the following fermentation conditions described in EP 177882, the factors in which R ′ ₂ is (C ₁₀ -C ₁₁ ) alkyl are very significant. though it is, R _'2 is C ₉ or C
It is possible to increase the amount of minor constituents that are ₁₂ alkyls by changing the fermentation conditions.

During the normal purification steps of the antibiotic A40926 complex, factors PA and PB are largely converted to factors A and B.

In addition, the antibiotic A40926 complex, a single factor thereof or a mixture of said factors, in all proportions, is subjected to limited acid hydrolysis of one of the sugar moieties of the starting material to yield the corresponding N-acylaminoglucuronyl aglycone. It was found that the complex AB, N-acylaminoglucuronyl aglycone factor A, N-acylaminoglucuronyl aglycone factor B, and A40926 can be converted into mannosyl aglycone (European Patent Application Publication No. 240609 and European Patent Application). Published No. 228015, see).

Suitable hydrolysis conditions for the formation of N-acylaminoglucuronyl aglycone include the use of dimethylsulfoxide / concentrated hydrochloric acid 8: 2-9.5-0.5 at a temperature of 40-80 ° C.

The antibiotic A40926 N-acylaminoglucuronyl aglycone has R ′ ₁ and M ′ are hydrogen atoms, X ′ is hydroxy, Y ′ is carboxy, and R ′ ₂ is (C ₉ -C
₁₂ ) Represented by formula (II) above when alkyl.

Complete cleavage of the entire sugar moiety of antibiotic A40926 gives the aglycone. This hydrolysis method is described in European Patent Application Publication No. 24
No. 0609.

The antibiotic A40926 complex, its factors, their corresponding N-acylaminoglucuronyl aglycones, mannosyl aglycones, aglycones, and their mixtures in all proportions are mainly antibacterial against Gram-positive bacteria and Neisseriae. Have activity.

In International Patent Application No. PCT / EP92 / 00374 claiming priority of European Patent Application No. 91104857, antibiotic A40926 and its ester derivatives of N-acyl-aminoglucuronyl aglycone (position 6 ^B , ie N- Esterification at the carboxy group present on the acylaminoglucuronyl moiety) is described; for example, X ′ is OH,
Y 'is (C ₁ -C ₄₎ alkoxycarbonyl and R,' _1,
Compounds of formula (II) when R _'2 and M' have the same meanings as the symbols R _1, R ₂ and M.

These ester derivatives are N ¹⁵ protected (in this description, the term “N ¹⁵ ” means A 4 which is appropriately given the number 15.
Reacting a nitrogen atom of an amino functional group attached to a carbon atom of a molecule) or N ¹⁵ free amino A40926 substrate or its demannosyl derivative (ie, N-acylaminoglucuronyl aglycone) with an alkanol in an acidic medium. Alternatively, a N ¹⁵ protected A40926 derivative or a demannosyl homolog thereof and a halogenated (preferably
Alkyl bromide, chloride or iodide), optionally in the presence of a hydrohalic acid acceptor, especially in excess of the selected alkanol in the presence of concentrated mineral acid, at temperatures between 0 ° C and room temperature. , Made by reacting.

These ester derivatives of antibiotic A40926 prepared by the above method are used as starting materials for the preparation of antibiotic A40926 derivatives of formula (I).

As outlined above, the starting materials for the compounds of the invention are A40926 ester derivatives and demannosyl A409.
A limited esterification method useful for making 26 ester derivatives is an excess of A40926 substrate in the presence of concentrated mineral acid at temperatures between 0 ° C. and room temperature during which the A40926 substrate changes the steric complexity of the group to be introduced. Includes an esterification reaction that is mixed with the selected alkanol.

In some instances, it is convenient to protect the primary amino functionality at position 15 of the A40926 precursor to reduce possible unwanted side reactions. this is,
Performed by methods known per se in the art, as described in the reference books below (see TW Green,
"Protective Groups in Organic Synthesis", John Wil
ey and Sons, New York, 1981 and M. Mc Omie, “Protect
ive Groups in Organic chemistry ", Plenum Press, New
York, 1973). These protecting groups must be stable under the conditions of the reaction process, they do not interfere with the main reaction, and must be easily cleaved at the end of the main reaction.

Tert-butoxycarbonyl (t-BOC), carbobenzyloxy (CBz), and arylalkyl groups are examples of suitable amino protecting groups. Benzylation with optionally substituted benzyl halides in the presence of base readily occurs in quantitative yield, yielding exclusively the corresponding N ¹⁵ -benzyl derivative without concomitant formation of the benzyl ester of the carboxy group. Bring formation.

Selective protection of the amino group at position 15 is preferred by reaction with benzyl bromide in the presence of a hydrogen halide acceptor (ie a tertiary amine) without concomitant esterification of the two carboxy groups. It can be carried out.

The conditions for removal of the N ¹⁵ -protecting group are methods known in the art for the removal of amino protecting groups and must be determined after considering the reactivity of other free radicals in the molecule.

M'is α-D-mannopyranosyl or 6-O-acetyl-α-D-mannopyranosyl, and Y'is (C _1-
The ester starting material of formula (II) when C ₄ ) alkoxycarbonyl is converted to the corresponding compound when M ′ is hydrogen by the method of selective acid hydrolysis. Suitable hydrolysis conditions for making demannosyl derivatives of antibiotic A40926 (eg N-acylaminoglucuronyl aglycone), as disclosed in EP-A-240609, are temperatures of 40-80 ° C. In dimethyl sulfoxide / concentrated hydrochloric acid 8: 2 (v / v) to 9.5: 0.5 (v / v).

Thus, demannosyl derivatives of the ester of A40926 are obtained in the form of a mixture with the corresponding aglycones and can be fractionated by preparative HPLC. The hydrolysis conditions can also be modified as appropriate to change the ratio between the products obtained. For example, starting from A40926 esterified at the 6 ^B position, by increasing the solvent / hydrochloric acid ratio to 78: 1, maintaining the reaction temperature below 60 ° C., and extending the reaction time to about 7 days, A40926 The ratio of the required demannosyl derivatives esterified at position 6 ^B to the unwanted aglycones of is about 1.4-1.0.

Reaction progress is monitored by HPLC according to methods known in the art. On the basis of the results of these measurements, the person skilled in the art evaluates the progress of the reaction, includes the termination of the reaction and, for example, solvent extraction, non-solvent precipitation and, in combination with chromatographic fractionation and purification, essentially It would be possible to determine when to start collecting the reactants according to known techniques.

The ester derivatives used as starting materials for the preparation of compounds of formula (I) are the precursor antibiotic A40926
It may be a single compound corresponding to each of several factors of the complex or a mixture of two or more components corresponding to different factors of the A40926 precursor in any ratio. The mixture of ester derivatives may be derived from the use of the A40926 complex or a factor mixture of A40926 complex precursors in the production of a 6 ^B ester, or the resulting ester product (the first of the factors identifying the precursor A40926 complex). Pure ester, which can be varied in proportion) by the application of special conditions in the isolation / purification, or by the reverse phase chromatographic fractionation method or by using the pure A40926 factor as a precursor It can be obtained by mixing the products in suitable proportions.

In this specification and in the claims, unless otherwise specified, the term "alkyl", whether alone or in combination with other substituents, includes both straight-chain and branched hydrocarbon radicals; more particularly , The term “(C ₁ -C ₄ ) alkyl” means 1 to 1 carbon atoms such as methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 1,1-dimethylethyl and 2-methylpropyl. Represents 4 straight or branched aliphatic hydrocarbon chains.

As used herein, the terms “alk ₁ ”, “al
"k ₂ ", "alk ₃ " represents an independent linear or branched alkylene chain of 2 to 10 carbon atoms, such as: The term as used herein, "(C ₂ -C ₄₎ alkyl moieties" and "(C ₂ -C ₄₎ alkylene moiety" 2-4 linear or branched aliphatic chain carbon atoms Represents a residue. Representative examples of the chains can be cited from the list above.

The term “(C ₁ -C ₄ ) alkoxycarbonyl” refers, for example, to methoxycarbonyl, ethoxycarbonyl, propyloxycarbonyl, isopropyloxycarbonyl,
Butoxycarbonyl, isobutoxycarbonyl, and
Includes both straight chain and branched alkoxycarbonyl groups such as tert-butoxycarbonyl.

Hereinafter, the definition amino acid residue by -NR ₃ -alk ₁ - show some typical examples of _{(NR 5 -alk 3) q -W} - (NR 4 -alk 2) p: And the like.

If R ₃ and R ₄ together represent a (C ₂ -C ₄ ) alkylene moiety bound to two nitrogen atoms, that moiety is
The saturated heterocyclic moiety formed in the combination of alk ₁ (or alk ₂ ) and two adjacent nitrogen atoms is preferably a piperazino ring.

For example, when R ₃ and R ₄ (or R ₄ and R ₅ ) together represent a (C ₂ -C ₄ ) alkylene moiety bonded to two nitrogen atoms, or for p and q When both are zero and W together with the moiety -NR ₃ -alk _1- represents piperazino or 4-methylpiperazino, the formula -NR ₃ -alk _1- (NR ₄ -alk ₂ ) p- ( amino acid residues of NR ₅ -alk ₃₎ q-W is the same as the following groups: The scope of the present invention is derived from a unit compound of formula (I) obtained from a single factor of precursor antibiotic A40926 complex and also from complex A40926 itself or a mixture of two or more of the factors in any proportion. Included are mixtures of compounds of formula (I) Therefore, A409
The change in the mutual proportions of the mixture components of the compound of formula (I) corresponding to the factor of 26 complex was determined by applying various conditions in fermentation, recovery, isolation and purification conditions of precursor antibiotic A40926 complex. Or by mixing each of the isolated factors of the esters of formula (II) in the desired proportions and then converting them into compounds of formula (I), or of the compounds of formula (I) of the present invention. It can be obtained by mixing the pure individual factors in the desired proportions.

Suitable compounds of formula (I) are: and their pharmaceutically acceptable addition salts.

  In this case, R₁Is a protective group for hydrogen or amino functional groups
Represents; R₂Is (C₉-C₁₂) Represents alkyl; M is hydrogen,
α-D-mannopyranosyl or 6-O-acetyl-
represents α-D-mannopyranosyl; Y is carboxy,
(C₁-C_Four) Alkoxycarbonyl, aminocarbonyl,
(C₁-C_Four) Alkylaminocarbonyl, di (C₁-C_Four)
Represents alkylaminocarbonyl, the alkyl part of which
Minutes are hydroxy, amino, (C₁-C_Four) Alkylamino
And J (C₁-C_Four) One selected from alkylamino
It may have a substituent or hydroxymethyl; X is
Hydroxy or formula -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_FourAnd R_FiveRepresents hydrogen; alk₁, Alk₂Oh
And alk₃Are each independently linear with 2 to 4 carbon atoms.
Represents a branched alkylene; p and q are independently 0 or
Is an integer representing 1; W is hydrogen, (C₁-C_Four) Al
Kill, amino, (C₁-C_Four) Alkylamino, di (C₁−
C_Four) Alkylamino, 1 or 2 amino- (C₂−
C_Four) Alkyl moieties or 1 or 2 (C₁-C_Four)
Rukiruamino- (C₂-C_Four) Alkyl moiety or 1
Is two di (C₁-C_Four) Alkylamino- (C₂-C_Four) Al
Represents an amino substituted by a kill moiety, or
If p and q are both zero, then W is a partial −NR
₃−alk₁-With, again, piperazino or
May represent 4-methylpiperazino] In the case where X represents hydroxy,
In, Y represents hydroxymethyl, Z is hydrogen or
Is based [In the formula, A Is an anion of an inorganic or organic acid, or
Is a carboxylic acid functional group on the remaining part of the antibiotic.
If present, it is also derived from the carboxylic acid functionality.
May represent internal anions that are induced] Represents

Other suitable compounds of the invention are those wherein R ₂ is (C _10-
C ₁₁ ) alkyl, M represents α-D-mannopyranosyl, and R ₁ , X, Y and Z are as defined above, including derivatives of formula (I) and pharmaceutically acceptable addition salts thereof. To do. More preferred compounds of the present invention are
Includes the following cases of compounds of formula (I) as well as their pharmaceutically acceptable addition salts.

In this case, R ₁ represents hydrogen or a protecting group for an amino functional group, preferably hydrogen; R ₂ represents 7-methyloctyl, n-nonyl, 8-methylnonyl, n-decyl, 9-
Methyldecyl, n-undecyl or n-dodecyl,
Preferably n-decyl, 9-methyldecyl or n-
Represents undecyl, most preferably 9-methyldecyl; M
Represents hydrogen or α-D-mannopyranosyl, preferably α-D-mannopyranosyl; Y represents carboxy, (C ₁ -C ₄ ) alkoxycarbonyl, aminocarbonyl, (C ₁ -C ₄ ) alkylaminocarbonyl, Di (C ₁ −
C ₄ ) alkylaminocarbonyl, wherein the alkyl moiety is one substituent selected from hydroxy, amino, (C ₁ -C ₄ ) alkylamino and di (C ₁ -C ₄ ) alkylamino or hydroxy. Methyl, preferably carboxy, methoxycarbonyl, aminocarbonyl, methylaminocarbonyl, dimethylaminocarbonyl,
(Dimethylamino) may have ethylaminocarbonyl or hydroxymethyl; X is an amino acid residue _{-NR 3 -alk 1 - (NH-} alk 2) p- (NH-alk 3) q-W [ wherein , R ₃ represents hydrogen; alk ₁ , alk ₂ and alk ₃ are
Each independently represents a linear alkylene having 2 to 4 carbon atoms; p and q each independently represent 0 or 1, and W is amino, (C ₁ -C ₄ ) alkylamino, di ( C ₁
-C ₄ ) alkylamino, 1 or 2 amino- (C ₂
-C ₄₎ an amino substituted by alkyl moiety,
Alternatively, when p and q are both zero, W is taken together with the moiety —NR ₃ —alk ₁ — and also represents piperazino or 4-methylpiperazino], most preferably X is —NH _{- (CH 2) 3 -N (} CH 3) 2, -NH- (CH 2) 3 - [NH (CH 2) 3] 2 -NH 2, -NH- (CH 2) 3 -N [(CH 2 ) ₃ NH ₂ ] ₂ and Is an amino residue selected from and Z represents hydrogen.

Y is (C ₁ -C ₄ ) alkoxycarbonyl, R ₁ ,
R _2, M and Z are as were clearly at the beginning of this specification, and X is an amino acid residue _{-NR 3 -alk 1 - (NR 4} -alk 2) p- (NR 5 -alk 3) q -W [wherein R ₃ , R ₄ , R ₅ , alk ₁ , alk ₂ , alk ₃ , p, q, and W are as defined at the beginning of the present specification] (I compounds of) is, R _'1, R' ₂ and M '
Have the same meanings as R ₁ , R ₂ and M, X ′ is hydroxy and Y ′ is (C ₁ -C ₄ ) alkoxycarbonyl, the corresponding derivatives of formula (II) above. Produced by amidation of.

These starting materials of formula (II) were prepared as described above, one particular example of which is the already mentioned International Patent Application No. PC
It is disclosed in T / EP92 / 00374.

The amidation method is performed by using the starting material of the formula (II) and the formula (II
I): Where R ₃ , R ₄ , R ₅ , alk ₁ , alk ₂ , alk ₃ , p, q and W have the same meanings as defined at the beginning of the specification. In the presence of an agent or the formula (I
I) via formation of the "active ester" of the starting C ⁶³ carboxylic acid, followed by condensation in an inert organic solvent.

Inert organic solvents useful for the amidation reaction are organic aprotic solvents that can solubilize the starting material at least in part without adversely interfering with the reaction progress.

Examples of said inert organic solvents are organic amides, ethers of glycols and polyols, phosphoric acid amides and sulfoxides. More preferred examples of inert organic solvents are:
Dimethylformamide, dimethoxyethane, hexamethylphosphoramide, dimethylsulfoxide and mixtures thereof.

The condensing agent in the method of the present invention is suitable for forming an amide bond in organic compounds and particularly in peptide synthesis.

Representative examples of condensing agents are diisopropylcarbodiimide (DIC), dicyclohexylcarbodiimide (DCC) in the presence of hydroxybenzotriazole (HBT),
Benzotriazolyloxy-tris- (dimethylamino) phosphonium hexafluorophosphate, Benzotriazolyloxy-tris- (pyrrolidino) phosphonium hexafluorophosphate, and diphenyl azidoate, diethyl azidolate, di- (4) - nitrophenyl), such as Ajidorin acid Jimoruhoriru (C ₁ -
C ₄ ) alkyl, phenyl or heterocyclic azido acid esters and diphenyl chlorophosphate. Suitable condensing agents are diphenyl azidophosphate, i.e. phosphoric acid diphenyl ester azide (DPPA), hexafluorophosphoric acid benzotriazolyloxy-tris- (dimethylamino) phosphonium (BOP), and hexafluorophosphoric acid benzotriazolyl. Oxy-tris- (pyrrolidino)
Phosphonium (PyBOP).

Of the last two condensing agents mentioned above, PyBOP is particularly preferred because the resulting by-product pyrrolidine is less toxic than dimethylamine.

In the amidation process of the invention described herein, the reaction in some cases involves equimolar proportions, or a slight molar excess of the amine reactant, especially when BOP or PyBOP is used as the condensing agent. Although usually performed in high yield, the amine reactant is usually used in molar excess.

Generally, where the amine reactant is a fairly inexpensive or readily available reactant, a 2-10 fold molar excess of amine (III) is used, but a 3-4 fold molar excess is more preferred. .

In order to carry out the amidation of the starting material of the above formula (II) with amine (III) in the presence of a condensing agent, the amine reactant is linked to the carboxy functional group (X ′ = hydroxy) of the starting material. It is necessary to be able to form salts. If the amine is unable to form such a salt sufficiently strongly in the chosen reaction medium, a salt-forming base (for example, a tertiary aliphatic such as triethylamine, N-methylpyrrolidine or N-methylpiperazine or Heterocyclic amines, which cannot form amide bonds with carboxy functional groups)
It is necessary to add to the reaction mixture in at least equimolar amounts with respect to the starting materials.

Using a lower molar excess of the amine reactant by addition of a salt forming base is the preferred method when the amine reactant is relatively expensive or difficult to obtain.

Examples of such salt-forming bases are tertiary aliphatic or heterocyclic amines such as trimethylamine, triethylamine, N-methylpyrrolidine or picoline, and the like.

  The condensing agent is generally an equimolar amount or 1.1 to 1.7 times, preferably
The starting material should be in a slightly excess molar amount such as 1.2 to 1.5 times.
Used for quality A40926 compounds. Especially as a condensing agent
Large excess (eg, 3-fold molar excess) of PyBOP and large
Excess (eg, 6-10 fold molar excess) of amine reactant
If used, Y'is (C₁-C_Four) Alkoxycarbonyl
Depending on the starting material of formula (II) where Z is [In the formula, A Has the same meaning as above] The amide-terminated product of formula (I) when
It was observed to be obtained in a reasonable yield.

The amine reactant may also be conveniently added to the reaction medium as the corresponding acid addition salt, eg the hydrochloride salt. In this case, a strong base capable of liberating the amine from its salt, at least twice the molar amount, preferably 2-4.
A double molar excess is added. Also in this case, suitable bases are usually tertiary organic aliphatic or heterocyclic amines which are incapable of forming amide bonds with the carboxy functional groups as exemplified above. In fact, in at least some instances it is highly preferred to use salts of the amine that are dissociated in situ by the base, especially if the salt is more stable than the corresponding free amine. .

The reaction temperature will vary considerably depending on the particular starting materials and reaction conditions. Generally, it is preferable to carry out the reaction at a temperature of 0 to 30 ° C.

Also, the reaction time will vary considerably depending on the condensing agent and other reaction parameters. Generally, the condensation reaction is completed within about 1 hour to about 24-48 hours.

In each case, the progress of the reaction is monitored by TLC or preferably HPLC according to methods known in the art.

Based on the results of these analyses, the person skilled in the art evaluates the progress of the reaction, includes the termination of the reaction and, for example, solvent extraction, precipitation by addition of a non-solvent, etc. It would be possible to determine when to begin recovering the reactants according to techniques known per se, which are carried out in combination with chromatographic purification.

Commonly, when using a condensing agent as described above, N ¹⁵ of the starting esters of formula (II) - to protect the amino function is not necessarily required. However, where the ester is made from the precursor antibiotic A40926,
Utilizing a starting ester protected at such a functional group if it results directly from the preceding reaction step is
May be beneficial. Furthermore, there may be special cases where the amidation reaction conditions require, or at least make more suitable, the N ¹⁵ -amino function in the starting ester of formula (II). Absent.

In such a case, the N ¹⁵ -amino functional group is protected against protection of the A40926 precursor for the formation of an ester of formula (II) [where Y ′ is (C ₁ -C ₄ ) alkoxycarbonyl]. And protected by methods known per se in the art, as described in the previously referenced reference books.

The N-protecting group must be stable under the conditions of the reaction, must not interfere adversely with the amidation reaction, and the newly formed amide bond as well as the overall structure of the compound, eg Easily separated from the reaction medium at the end of the reaction without changing the sugar moiety,
Must be removed.

In order to protect the N ¹⁵ -primary amino function of the ester starting material, as well as all other amino functions optionally identified as amine (III) that do not participate in the amidation reaction, if necessary, Representative of N-protecting groups which may be advantageously used in the process of the present invention are carbamate-forming reagents characterized by the following oxycarbonyl groups: 1,1-dimethylpropynyloxycarbonyl,
t-butyloxycarbonyl, vinyloxycarbonyl, cinnamyloxycarbonyl, benzyloxycarbonyl, P-nitrobenzyloxycarbonyl, 3,4-dimethoxy-6-nitrobenzyloxycarbonyl, 2,4
-Dichlorobenzyloxycarbonyl, 5-benzisoxazolylmethyloxycarbonyl, 9-anthranylmethyloxycarbonyl, diphenylmethyloxycarbonyl, isonicotinoyloxycarbonyl, S-benzyloxycarbonyl, and the like.

Generally, these protecting groups can be removed at the end of the amidation reaction by treatment with a suitable strong organic acid such as trifluoroacetic acid (TFA) and a dilute inorganic acid.

With protecting groups to avoid the risk of hydrolysing the sugar moiety at the core of the antibiotic molecule and under different removal conditions, such as catalytic hydrogenation using palladium on carbon as a catalyst. Can also be detached. On the other hand, it is also possible to eliminate amino protecting groups selected from those mentioned above under controlled acidic conditions, for example at low temperatures and / or short reaction times.

When the amidation reaction is carried out through intermediate formation of the "active ester" of the starting compound of formula (II),
Such "active esters" are generally either formed as such, or else may be isolated and then reacted with an amine of formula (III). The starting materials of formula (II), preferably, N ¹⁵ - in order to avoid any interference with the active ester formation reagent by amino groups, N ¹⁵ - is protected at the amino functional group. Protection of such groups is accomplished according to known methods and those described above.

The formation of "active esters" of carboxylic acids is described by "Fieser a
nd Fieser, Reagent for organic synthesis, John Wiley
and Sons Inc., pages 129-130 (1967) ”,
It is written in general terms.

Examples of such active ester-forming reagents that can be conveniently used in the method of the invention are "R. Schwyzer et al., Helv. Chim.
Acta, 38 , 69-70 ", wherein ester derivatives of formula (II) [wherein X'is CH ₂ CN, CH ₂ COOC ₂ H
₅ , CH ₂ (COOC ₂ H ₅ ) ₂ , CH ₂ COCH ₃ , Is a starting material of formula (II) [wherein R ′ is
₁ is a suitable protecting group and X'is hydroxy]
From ClCH ₂ C in the presence of acid acceptor in the solvent, respectively.
N, BrCH ₂ COOC ₂ H ₅ , BrCH (COOC ₂ H ₅ ) ₂ , ClCH ₂ COCH ₃ , Can be produced by reacting with.

A suitable reagent of this type is chloroacetonitrile. In this case, chloroacetonitrile itself, dimethylformamide (DMF) or dimethylsulfoxide (DMSO) can be used as a suitable solvent.

In general, inert organic solvents useful for forming "active esters" are organic aprotic solvents that can dissolve the carboxylic acid starting material, at least in part, without adversely interfering with the progress of the reaction.

Examples of said inert organic solvents include organic amides, ethers of glycols and polyols, phosphoramides,
These are sulfoxides and aromatic compounds. Suitable examples of inert organic solvents are: dimethylformamide, dimethoxyethane, hexamethylphosphoramide, dimethylsulfoxide, benzene, toluene and mixtures thereof.

More preferably, the solvent is selected from acetonitrile, dimethylsulfoxide, dimethylformamide. The formation of active esters generally does not interfere with the progress of the reaction, trialkylamines such as triethylamine, carbonic acid or sodium or potassium bicarbonate,
In the presence of a base such as Generally, the base is used in a 2 to 6 times molar proportion relative to the starting material,
Preferably, it is used in about a 3-fold molar excess. The preferred base is triethylamine.

The “active ester” forming reagent is used in large excess relative to the C ⁶³ carboxylic acid starting material of formula (II). In general,
It is used in a proportion of 5 to 35 mol, preferably in about a 20 to 30 fold molar excess. The reaction temperature is 10 to 60 ° C, preferably 15 to 30 ° C. Usually, the reaction time depends on the other specific reaction parameters and can generally vary between 3 and 48 hours.

The progress of the reaction is followed by HPLC or TLC,
An operation is initiated to determine when the reaction is considered complete and to recover the desired intermediate. The "active ester" intermediate is used directly in the same reaction medium in which it is produced, but is generally isolated by precipitation without solvent or by solvent extraction for further purification. Instead, it is used as such for the next reaction step. However, if desired, it can be purified by column chromatography, such as flash column chromatography or reverse phase column chromatography. The resulting “active ester” intermediate is then reacted with a molar excess of the amine derivative of formula (III) in the presence of an organic polar solvent at a temperature of 5-60 ° C., preferably 10-30 ° C. .

The organic polar solvent is in this case a polar protic solvent or an aprotic solvent.

Suitable examples of organic polar protic solvents include ethanol, n
A lower (C ₂ -C ₄ ) alkanol, such as -propanol, iso-propanol, n-butanol and the like, or mixtures thereof, preferably
Used in anhydrous form.

Suitable examples of the organic polar aprotic solvent include N, N-dimethylformamide (DMF), hexamethylphosphoramide (HMPA), or a mixture thereof, 1,3-dimethyl-
3,4,5,6-Tetrahydro-2 (1H) -pyrimidone (DMP
U), dimethyl sulfoxide (DMSO) or dimethoxyethane (DME).

The reaction of the selected amine of formula (III) with the "active ester" can be carried out at temperatures of 5-60 ° C, although suitable temperatures are
Generally 10-30 ° C, most preferably 20-25 ° C,
Furthermore, the preferred molar ratio of the "active ester" intermediate to the amine (III) as defined above is 1: 5 to 1:30, more preferably 1:10 to 1:20. The reaction progresses as usual in TL
Monitored by C or HPLC.

When the reactant amine is a polyamine of formula (III), one or more amino groups without formation of an amide bond may be conveniently protected. Again, in these cases, suitable protecting groups are those already mentioned for N ¹⁵ .

Thus, the resulting N ⁶³ -protected amide derivative is then deprotected under conditions similar to those described above for the 15-position deprotection.

Formula (I) wherein Y is hydroxymethyl and R ₁ , R ₂ , M, X and Z are as described at the beginning of the specification.
Compounds of formula (I) wherein R ₂ , M, X and Z have the same meanings as above, Y is (C ₁ -C ₄ ) alkoxycarbonyl,
And the corresponding derivatives of formula (I) when R ₁ is a suitable protecting group for the N ¹⁵ -amino function, preferably selected from sodium borohydride, potassium borohydride and sodium cyanoborohydride. Formed by reaction with an alkali metal borohydride at a temperature of 0 to 40 ° C. in an aqueous or alcoholic aqueous liquid.

The use of this method is such that Y is hydroxymethyl and X is
Is hydroxy, R ₁ , R ₂ , and M are as described at the beginning of this specification, and is specifically required to produce compounds of formula (I) where Z is hydrogen. It In this case, the starting material that is subjected to the reduction step under the above conditions is that Y ′ is (C ₁ -C ₄ ) alkoxycarbonyl, X ′ is hydroxy and R ′ ₂ and M ′ are each R ₂ And M having the same meanings and R ′ ₁ is a suitable protecting group for the N ¹⁵ -amino function, a compound of formula (II). The special production of the starting material is described in International Patent Application No. PC.
T / EP92 / 00374, which has the formula (I
It is carried out according to the general procedure for producing the starting ester of I).

Generally, the hydroalcoholic liquid used in the above reduction reaction is a mixture of water and a lower alkanol which is water-soluble or partially mixed with water, and the water / lower alkanol ratio thereof is 40/60 to 90/10. (V / v), preferably 60/40 (v /
v) to 68/32 (v / v), most preferably 65/35 (v /
v).

The reaction may, in some cases, also be in the presence of relatively small amounts of water, eg water / lower alkanol 30/70 or 20/80.
In general, the reaction rate is very low when the water / lower alkanol ratio is 40/60 or less, although it occurs in the mixed solution.

Suitable lower alkanols include linear and branched (C _1-
C ₄₎ alkyl alcohols, the best ones, n- butanol, ethanol and methanol. Sometimes, in special cases, polar co-solvents such as N, N-dimethylformamide, 1,3-dimethyl-3,4,5,6-tetrahydro-2 (H1) -pyrimidone (DMPU) and dimethylsulfoxide. Can be added to completely dissolve the starting materials during the course of the reaction. Sometimes also various amounts of diethyl ether are added to avoid foaming.

Sodium borohydride is the most suitable alkali metal borohydride. The appropriate amount of alkali metal borohydride provided varies depending on the solvent used as well as the reaction temperature, but is such that the pH of the reaction mixture is neutral or alkaline, preferably pH 7-10. It is desirable to use an amount of alkali metal borohydride in the process in a large excess over the stoichiometrically required amount. Generally, the molar ratio of alkali metal borohydride and its antibiotic starting material is between 50 and 300.

The reaction temperature can vary considerably depending on the particular starting materials and reaction conditions. Generally, it is preferred to react at 0-40 ° C, more preferably at room temperature.

The reaction time can also be varied considerably depending on other reaction parameters but must be carefully controlled. Generally, the reaction is complete in about 1-4 hours. If the reaction is extended for more than 4 hours, undesired side reactions can occur that can cause the cleavage of the core peptide bonds of the molecule.

In all cases, reaction progress is monitored by TLC by methods known in the art or, preferably, by HPLC. Based on the results of these measurements, a person skilled in the art evaluates the progress of the reaction, includes stopping the reaction and, for example, solvent extraction, precipitation by addition of a specific solvent, etc. It will be possible to determine when to begin the recovery of the reactants according to techniques known per se.

After the reaction was completed, excess alkali metal borohydride, for example, (C ₁ -C ₄₎ polar protic solvent dissolved acids such as alkyl alcohols, for example, (C ₁ -
C ₄₎ alkyl organic acid, (C ₁ -C ₆₎ alkyl sulfonic acid,
It is removed by adding the appropriate amount of aryl sulfonic acid and the like.

In addition, Y is hydroxymethyl, R ₁ , R ₂ ,
And M are as described at the beginning of the specification,
X is an amino acid residue _{-NR 3 -alk 1 - (NR 4} -alk 2) p- (NR 5 -alk 3) q-W [ _{wherein, R 3, R 4, R} 5, alk 1, alk 2, alk ₃ , p, q and W have the meanings given at the beginning of the specification] and Z is hydrogen, the compounds of formula (I) are compounds wherein Y is hydroxymethyl and X is hydroxy. And R ₁ , R ₂
And M has the same meanings as above, and Z is hydrogen, the corresponding compound of formula (I) is
Is produced by following an amidation procedure similar to that described above by reacting with an amine of.

Further, in this case, the amidation reaction, the "active ester for the production of suitable either by use of a condensing agent, or Y is (C ₁ -C ₄₎ compounds of formula (I) alkoxycarbonyl It is carried out through the formation of the intermediate.

In general, amidation of derivatives of formula (I) where Y is hydroxymethyl, X is hydroxy, and Z is hydrogen using PyBOP as the condensing agent, the PyBOP is in a large molar excess relative to the carboxylic acid starting material. Even when offered to
The final compound of formula (I) is produced when Z represents hydrogen. When the amidation reaction is carried out via the formation of an "active ester" of a compound of formula (I) when X is hydroxy, Y is hydroxymethyl and Z is hydrogen, it is It is preferred that the N ¹⁵ -amino group of the compound is protected.

Y is (C ₁ -C ₄ ) alkoxycarbonyl or hydroxymethyl, R ₁ , R ₂ , M and Z are as indicated at the beginning of the specification and X is an amino residue —NR ₃ -alk. _{1 - (NR 4 -alk 2)} p- (NR 5 -alk 3) q-W [ _{wherein, R 3, R 4, R} 5 are hydrogen or each independently (C ₁ -C
₄ ) represents alkyl, alk ₁ , alk ₂ , alk ₃ and W are as indicated at the beginning of the specification, p is 1 and q represents 1 or zero. A further method for the production of compounds of I) is that Y, R ₂ , M and Z are as described above and X is an amino residue —NR ₃ —alk ₁ —NHR _{4 where} R ₃ , R ₄ and alk ₁ are as described above] or —NR ₃ —alk ₁ —NR ₄ —alk ₂ —NHR ₅ [wherein R ₃ , R ₄ , R ₅ , alk ₁ and alk _{2 are} Is as described above] N ⁶³ amide of formula (I) (in this specification,
The term “N ⁶³ ” refers to the N ¹⁵ -protected derivative of the carboxamide group containing the carbon atom of the A40926 molecule identified by the numeral 63) and a formula (IV) or (IV
a), respectively [Wherein R ₅ , alk ₂ and alk ₃ and W are as described above, q is zero or 1 and r represents halo, methanesulfonyl or tosyl]. Consisting of reacting in the presence of an acid acceptor in an active solvent.

The N ¹⁵ -protected derivatives of N ⁶³ amide described above are produced by the general method for the production of compounds of formula (I) of the present invention. N ¹⁵ - deprotection of the amino function is carried out by the condition.

Further, in the case of the above alkylation method, N ^{63 of the} formula (I) is used.
It may be useful or necessary to protect these amino functional groups of the amine reactant (IV) or (IV a) other than the N ¹⁵ -amino group of the amide compound and / or not participating in the alkylation reaction. . The resulting ^N63 -protected amide can be deprotected under the above conditions.

The protecting groups used in all of the above reactions are those already mentioned. However, of particular note must be paid with respect to the deprotection step of the derivatives of formula (I) when Y is hydroxymethyl. For these compounds, in fact, when the 15-position protecting group is susceptible to elimination under acidic conditions, for example, dry trifluoroacetic acid (TF
The deprotection step is dangerous because there is a relatively fast antagonistic substitution of each 56-acylglucosamine moiety by treatment with A). However, separately, these undesired side reactions can easily be minimized. For example, t-butyloxycarbonyl (t-BO as a protecting group
If C) is used, the following conditions can be adopted: treatment with dry TFA at room temperature for 1 minute or at 0-5 ° C. for 10-30 minutes, followed by diethyl ether or methanol / 0-5 ° C. Rapid precipitation of the reaction product with a diethyl ether mixture. On the contrary, with the compounds of formula (I) when Y is carboxy or methoxycarbonyl, the 56-acylaminoglucuronic acid moiety is TF
It has been found to be significantly more stable to A. In fact, the corresponding minor formation of deglucuronyl pseudoaglycone is observed only after 1 hour reaction. However, in these cases t-BOC deprotection occurs in 30 minutes.

T with virtually no effect on the rest of the molecule
Other suitable methods for removing the -BOC protecting group include:
Consists of treatment with dichloromethane dissolved dry TFA at 0-10 ° C for 1-2 hours, followed by precipitation of reaction product by addition of non-solvent.

Compounds of formula (I) wherein R ₁ , R ₂ , M, X and Z are as set out at the beginning of the specification and Y is carboxy include compounds wherein Y is (C ₁ -C ₄ ) alkoxycarbonyl , Preferably methoxycarbonyl, and from all the corresponding compounds of formula (I) where all other symbols are as previously described, a temperature of 0 to 30 ° C. (higher than position 3 of the molecule). Must be avoided to prevent epimerization at the carbon atom of
Produced by treatment with an alkali metal hydroxide (eg, NaOH and KOH) in a di- (lower alkyl) ether of ethylene glycol or tetrahydrofuran. R ₁ , R ₂ , M, X and Z are as indicated at the beginning of the specification, Y is aminocarbonyl, (C _1-
C ₄ ) alkylaminocarbonyl, di (C ₁ -C ₄ ) alkylaminocarbonyl [wherein the alkyl moiety is hydroxy, amino, (C ₁ -C ₄ ) alkylamino and di (C ₁ -C ₄ ) alkyl. A compound of formula (I) which may have a substituent selected from amino] can also be produced according to the method shown below: i) The symbols Y and the partial COX of C ⁶³ are the same group ( C ₁ -C ₄ ) alkylaminocarbonyl or di (C ₁ -C ₄ ) alkylaminocarbonyl [wherein the alkyl moiety is amino, (C ₁ -C ₄ ) alkylamino and di (C ₁ -C ₄ )) It may have a substituent selected from alkylamino]
When (a) antibiotic A40926 complex, its demannosyl derivative or their factors (formula (II), X '= hydroxy, Y' = carboxy, R ₁ , R ₂ and M above) are represented. the same R of _'1, R' ₂ and M '), formula (II a
I) (wherein the symbols R ₃ , R ₄ , R ₅ , alk ₁ , alk ₂ , alk ₃ , p, q and W are carboxamide residues Y and CO as defined above).
Amidation with a large excess of the appropriate amine (having the meaning consistent with X). This amidation reaction is carried out under the same conditions as above.

ii) The partial COXs of the symbols Y and C ⁶³ represent different carboxamide residues, aminocarbonyl, (C ₁ -C ₄ ) alkylaminocarbonyl, di (C ₁ -C ₄ ) alkylaminocarbonyl [in which case the alkyl The moiety may have a substituent selected from hydroxy, amino, (C ₁ -C ₄ ) alkylamino and di (C ₁ -C ₄ ) alkylamino]
Generation of Derivatives When Representing the Meaning of Y Selected from X and the Meaning of X as an Amino Residue as Described at the Beginning of the Present Specification: Method A: R ₁ , R ₂ , M and Z are are as shown in the beginning of writing, X has the formula -NR ₃ -alk ₁ - in _{(NR 4 -alk 2) p- (} NR 5 -alk 3) q-W [ wherein all symbols are hereby As defined at the beginning of the text] and an amidation of the corresponding compound of formula (I) when Y is carboxy, a condensing agent under the same conditions as described above (eg , PyBOP or DPP
A) in the presence of A) with a suitable amine to form the carboxamide residue Y above; Method B: (a) protection of the same starting compound of Method A at the N ¹⁵ -amino functional group. (Eg, using a t-BOP or CBz group); (b) “active ester” formation of the carboxy group at position 6 ^B (eg, by reaction with chloroacetonitrile); (c) “active ester” of the compound.
Reaction of a moiety with a suitable amine to produce the carboxamide residue Y as defined above under the same conditions as above;
(D) N by optionally the method is carried out ¹⁵ - Removal of the protecting group (e.g., by acidolysis or hydrogenolysis).

Compounds of formula (I) in which M is hydrogen are generally prepared according to the above method using the corresponding starting molecule of formula (II) in which M ′ is hydrogen.

Furthermore, another method for the production of compounds of formula (I) in which M is hydrogen is by the method of selective acid hydrolysis according to the conditions described in EP-A-240609, wherein M is α
-D-mannopyranosyl or 6-O-acetyl-α
A compound of formula (I) which is -D-mannopyranosyl, M
Consisting of conversion to the corresponding compound where is hydrogen.

As mentioned above, the compounds of formula (I) consist of unit compounds which correspond to the individual factors of precursor antibiotic A40926 or mixtures thereof in different proportions. In most cases, the biological activity of the mixture is very similar to that of the individual factors, so there is no particular advantage in separating the individual components when obtaining the mixture. However, if the pure factors of formula (I) are required, they can be isolated from their mixture by reverse phase column chromatography according to the method described in EP 177882. Otherwise, they can be generated with unit starting materials of formula (II) corresponding to the individual factors of the antibiotic A40926 complex.

Under the general methods and conditions described here, one of the individual factors (eg factor B ₀ ) is
A particularly high proportion of the residual components of the mixture (eg,
It is beneficial to utilize the precursor antibiotic A40926 complex, which is contained at 60% in HPLC). Thus, a compound of formula (I) obtained from such a precursor through the process of the invention, unless otherwise subjected to the above separation method, is:
The particularly abundant components generally consist of mixtures corresponding to the same factors that are abundant in the A40926 complex precursor.

Methods for making the A-40926 complex enriched for its factors A and / or B ₀ , or PA and / or PB are described, for example, in EP-A-259781.

The compounds of the present invention possess a basic functional group which is capable of forming salts with organic and inorganic acids according to conventional methods.

Representative and suitable acid addition salts of compounds of the present invention include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, succinic acid, citric acid, ascorbic acid, lactic acid, maleic acid, Fumaric acid, palmitic acid, cholic acid, pamoic acid, mucus acid, glutamic acid, camphoric acid, glutaric acid, glycolic acid, phthalic acid, tartaric acid, lauric acid, stearic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, sorbic acid, Includes both organic and inorganic acids such as picric acid, benzoic acid, cinnamic acid, and the like, and salts formed by standard reactions.

The compounds of formula (I) in which X is hydroxy and Y is hydroxymethyl, as well as the compounds in which Y is carboxy, also have acidic functional groups that can form salts with organic and inorganic bases.

Representative examples of bases which can form salts with compounds of the invention having acidic functional groups are: sodium, potassium, calcium,
Magnesium, barium hydroxide, alkali metal or alkaline earth metal hydroxides such as ammonia, and aliphatic, cycloaliphatic or aromatic compounds such as methylamine, dimethylamine, triethylamine, ethanolamine and picoline. Organic amines.

Conversion of the "non-salt" compounds of the present invention to the corresponding addition salts and back conversion, eg conversion of the addition salts of the present invention to the non-salt form, are within the ordinary skill of the art and are encompassed by the present invention. It

For example, a compound of formula (I) is converted to the corresponding salt with an acid or base by dissolving or suspending the non-salt form in an aqueous solvent and adding a slight excess of the selected acid or base. be able to. The resulting solution or suspension is then lyophilized to recover the desired salt.

If the final salt is insoluble in the solution in which the non-salt form dissolves, the salt is in the non-salt form after addition of a stoichiometric amount or a slight excess of the selected acid or base. It may be recovered from the solution by filtration.

The non-salt form can be formed by neutralizing the corresponding salt dissolved in an aqueous solvent to liberate the non-salt form. Subsequently, it is recovered, for example by extraction with an organic solvent, converted to other addition salts by addition of the acid or base of choice and then harvested as described above.

If desalting is required following neutralization, the normal desalting procedure is used. For example, column chromatography on constant pore size polydextran resin (such as Sephadex LH20) or silanized silica gel is conveniently used. After elution of unwanted salts with an aqueous solution, the required product is a linear or stepwise gradient of water and polar or non-polar organic solvent, eg, acetonitrile / water 50:50 to about 100% acetonitrile. Is eluted with a solvent such as.

As is known in the art, salt formation with either pharmaceutically acceptable acids and bases or non-pharmaceutically acceptable acids and bases can be used as a convenient purification technique. After salt formation and isolation, the salt formation of the compound of formula (I) can be converted into the corresponding non-salt or pharmaceutically acceptable salt.

However, when dealing with the biological activity of the compounds of formula (I) in view of the similarity between the compounds of formula (I) and their salts, what is mentioned in this application is:
It also applies to their pharmaceutically acceptable salts.

The following Table I illustrates a series of representative compounds of the invention.

The antibiotic A40926 derivatives of the present invention are mainly active against Gram-positive bacteria.

In particular, the compounds of the present invention are useful for glycopeptide-resistant Enterococci and Staphylococci.
Against, it shows a surprising antibacterial activity.

The antibacterial activity of the antibiotic A40926 derivatives of formula (I) against selected strains of Gram-positive bacteria, expressed as the minimum inhibitory concentration (MIC), was determined in comparison with teicoplanin and antibiotic A40926 conjugates. Was done. Bovine albumin serum (fraction V / Sigma) 0.01% (w
/ v) containing Muller-Hinton
The microbroth dilution method in liquid medium was used. The final inoculum is about 10 ⁵ cfu / ml, MIC is 37 ° C., macroscopic growth is not observed minimum concentration after 18-24 hours incubation (mcg / m
read as l).

Table II below shows the antibacterial spectra of a series of representative compounds of the invention.

Table III, below, provides in vitro antibacterial activity data for some representative compounds of the present invention with respect to in vitro antibacterial activity against enterococcal strains highly resistant to glycopeptides in conventional therapy in comparison with teicoplanin and vancomycin. Shows.

Table IV below shows the results of some representative compounds of the invention against experimental streptococcal sepsis in mice.

This experiment is based on “V. Arioli et al., Journal of Antibio
tics 29 , 511 (1976) ”.

The above data show that the compound of the invention is a precursor A40926.
Than Neisseria gonorrhoe
Although the activity against ae) is generally low, it has been shown to have better antibacterial activity against clinical isolates of staphylococci and enterococci when compared to control compounds. In particular, they are: a) markedly higher in vitro antibacterial activity than teicoplanin and A40926 against staphylococci moderately or resistant to glycopeptides, especially methicillin-resistant staphylococci not producing coagulase; And in vitro antibacterial activity against highly glycopeptide-resistant enterococci with high resistance to vancomycin and some resistance to A40926 (MIC ≧ 64 mcg / ml); c) Teicoplanin and streptococcal sepsis in mice
It has a higher effect in vivo than A40926.

With regard to the above-mentioned antibacterial activity, the compounds of the present invention are useful for the prevention and treatment of infectious diseases caused by pathogenic bacteria sensitive to the active ingredient, in particular enterococci, streptococci and grapes which are insensitive to glycopeptide antibiotics It can be effectively used as an active ingredient of antibacterial formulations used in human medicine and veterinary medicine for the treatment of infections caused by coccus strains.

The compounds of the invention can be administered orally, topically or parenterally, with the parenteral route of administration being more preferred.

Depending on the route of administration, these compounds will be formulated into various dosage forms. Formulations for oral administration include capsules,
It may be in the form of tablets, solutions or suspensions. Capsules and tablets, in addition to the active ingredient, are well known in the art.
Diluents such as lactose, calcium phosphate, sorbitol and the like, lubricants such as magnesium stearate, talc, polyethylene glycol, binders such as polyvinylpyrrolidone, gelatin, sorbitol, tragacanth, acacia, flavoring agents , And conventional additives such as acceptable disintegrants and humectants. Liquid formulations are generally in the form of aqueous or oily solutions or suspensions and may contain conventional additives such as suspensions.

For topical use, the compounds of the invention may be formulated into formulations suitable for absorption through the mucous membranes or bronchial tissues of the nose and throat, and as a convenience, liquid propellants or inhalants, troches, Alternatively, it may take the form of a throat coating agent.

For administration to the eye or ear, the formulation may be in liquid or semi-liquid form. Topically applied agents are
It can be formulated in a hydrophobic or hydrophilic base such as an ointment, cream, lotion, coating or powder.

For anal administration, the compounds of the present invention are administered in the form of suppositories admixed with conventional vehicles such as cocoa butter, wax, spermaceti, or polyethylene glycol and their derivatives.

Injectable formulations include suspensions in oily or aqueous media,
It may take the form of a solution or emulsion and may contain formulation agents such as suspending, stabilizing and / or dispersing agents.

Alternatively, the active ingredient may be in powder form for reconstitution with a suitable solvent, such as sterile water, at the time of dosing.

The amount of active agent to be administered depends on various factors such as the size and condition of the subject being treated, the route and frequency of administration, and the causative agents involved.

The compounds of the present invention are generally effective at administration containing from about 1 to 40 mg of active ingredient per kg of body weight. Depending on the particular compound, infection and patient characteristics, effective doses will be administered in a single dose or in 2-4 divided doses per day. A particularly desirable formulation is about 30 per unit
Were produced in the form of dosage units containing about 500 mg.

Creating Example 1 starting material (MA) (Y 'is -COOCH _3, X' is -OH, R _'1 is -H, R' ₂ corresponds to the factor of the A40926 complex ( C ₉
-C ₁₂₎ alkyl, M 'is alpha-D-mannopyranosyl, antibiotic Z is obtained by a compound) European Patent Application Publication No. 177882 of Formula (II) where is -H
A40926 complex (150mg; 0.0866mmole) was added to methanol (3
0 ml) and the pH is adjusted to 2 with concentrated sulfuric acid.
The mixture is stirred at room temperature for 26 hours. A precipitate appears when the pH is brought to 6 with 0.15 ml triethylamine (TEA). After addition of diethyl ether, the precipitate is collected, washed thoroughly with diethyl ether and dried. Yield: 150 mg (99%).

Example 2 Creating compound 1 (RA) (Y is -CH ₂ OH, X is -OH, R ₁ is -H, R ₂ corresponds to the factor of the A40926 complex (C ₉ - C
₁₂ ) A compound of formula (I) when alkyl, M is α-D-mannopyranosyl and Z is —H) Step a: N ¹⁵ — (t-BOC) —MA Preparation According to Example 1 To a stirred solution of 1.8 g of the compound (MA) prepared and 1 g of sodium bicarbonate in 50 ml of a dioxane / water 1/1 solution is added a solution of 0.25 g of di-tert-butyl-dicarbonate in 5 ml of dioxane at 5 ° C. It is added dropwise over the course of a minute. After 1 hour at room temperature, the reaction mixture was
The pH is adjusted to 4 with 1N HCl. Then 150 ml of water are added and the resulting mixture is extracted with n-butanol (2 x 100 ml). The organic layer is separated and 100 ml of water
Washed at 40 ° C and a small amount (about 25
ml). The solid precipitated by addition of diethyl ether (100 ml) was collected and dried in vacuo overnight at room temperature to give the title compound pure enough for the next step.
^{N 15 - (t-BOC)} -MA1.6g is obtained.

Step ^{b: N 15 - (t-} BOC) to a stirred suspension of the compound 0.9g created by the step a creation water in 50ml of -RA, n-butanol / diethyl ether 1/1
30 ml of the mixture was added, followed by sodium borohydride
0.9g is added. The reducing agent is added in portions over 30 minutes at room temperature, then the reaction mixture is stirred at room temperature for 1 hour. After that, it is cooled down to 5 ℃, glacial acetic acid 1.5m
l is added, followed by 50 ml of water. The resulting mixture was extracted with n-butanol (100 ml) and the organic layer was collected as above and was sufficiently pure for the next final step c, the title compound N ¹⁵ −.
0.8 g of (t-BOC) -RA is obtained.

Step c: in dry trifluoroacetic acid (TFA) 5 ml, N ¹⁵ created by step b - (t-BOC) -RA0.5g solution, otherwise at room temperature for 1 minute (to, at 0 to 5 ° C., Stir (20-30 minutes) and then pour into 10 ml of a methanol / diethyl edel 1/4 mixture at 0-5 ° C. The title compound RA was collected by filtration, washed with diethyl ether and dried under vacuum at room temperature overnight to give the product 0.
Get 35g. A 0.15 g pure sample of compound RA is made by reverse phase column chromatography on silanized silica gel and collecting all fractions containing pure single component as described below.

Example 3: Compound 2 (MA-A-1 / B ₀ ) and Compound 6 (MA-A-
1) Preparation (Y is —COOCH ₃ and X is —NH— (CH ₂ ) ₃ —N (C
H ₃₎ is _2, R ₁ is -H, R ₂ is 9-methyldecyl (MA-A-1 / B 0) or A40926 corresponding to factor complex (C ₉ -C ₁₂₎ alkyl (MA -A-1),
M is alpha-D-mannopyranosyl, Z is a compound of formula (I) when it is -H) Method A Step ^{a: N 15 - (t-} BOC) -MA-A-1 of N being created DMSO30ml ^{To 1.5} g of a stirred solution of 15- (t-BOC) -MA (made by step a of Example 2 above) was added 3,3-
0.2 ml of dimethylamino-1-propylamine and 0.3 ml of diphenyl azidophosphate (DPPA) are added. After stirring for 4 hours at room temperature, 0.15 ml of DPPA is added and stirring is continued for another 20 hours at room temperature. 170m diethyl ether
solid is collected for precipitation by addition of l, the title compound N ¹⁵ - is (t-BOC) -MA-A -1 1.3g of the resulting.

Step b: The above product is dissolved in 10 ml TFA. The solution was stirred at room temperature for 20 minutes, then diethyl ether 90
ml is added. The precipitated solid was collected, washed twice with 50 ml of diethyl ether, and then dried under vacuum at room temperature overnight to obtain 0.9 g of the crude title compound (MA-A-1), which was purified by silanized silica gel. Reversed-phase chromatography on the column and combining only the fractions containing pure single factor of interest together to obtain pure MA-A-1 / B ₀
Is obtained.

Method B 1.8 g (about 1 mmol) of the compound of Example 1 (MA) in 30 ml DMF
In a stirred solution of 3,3-dimethylamino-1-propylamine 0.14 ml (about 1.15 mmol) and PyBOP 600 mg (about 1.2 m
mol) is added at room temperature. After stirring for 3 hours at 20-25 ° C,
150 ml of diethyl ether are added. The solid that precipitates is collected and then purified by reverse phase column chromatography and the fractions containing the pure single factor are combined together to give 1.15 g of compound MA-A-1.

Example 4: Creating a compound 7 (PyMA-A-1) (Y is -COOCH _3, X is _{-NH- (CH 2) 3 -N (} C
H ₃ ) ₂ , R ₁ is —H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, M
Is α-D-mannopyranosyl, and Z is P ⁺ (NC ₄ H ₈ ) ₃
Compound of formula (I) when CH ₃ COO ⁻ ) Compound MA1.8 prepared in Example 1 in 40 ml DMF
To a stirred solution of g (about 2 mmol), 3,3-dimethylamino-1
-Propylamine 2 ml (about 16 mmol) and PyBOP3.12 g
(About 6 mmol) is added at room temperature. After 30 minutes, the reaction mixture is purified as described in Example 3, Method B to give 1.5 g of the title compound PyMA-A-1.

Example 5 Compound 3 (RA-A-1 / B ₀ ) and Compound 8 (RA-A-
1) Preparation (Y is —CH ₂ OH and X is —NH— (CH ₂ ) ₃ —N (CH ₃ ).
₂ , R ₁ is —H, R ₂ is 9-methyldecyl (RA
-A-1 / B ₀ ) or (C ₉ -C ₁₂ ) alkyl (RA-A-1) corresponding to the factor of the A40926 complex, M is α-D-mannopyranosyl and Z is -H. a compound) method a step of formula (I) when ^{a: N 15 - (t-} BOC) -RA-a-1 creating following substantially example 3, method a, the same as described in step a the ^{method, N 15 - (t-BOC} ) -RA ( example 2, step b) from 2g, the title compound ^{N 15 - (t-BOC)} -
1.7 g of RA-A-1 is obtained.

Step b: The following essentially Example 2, by the same method as that described step c, the above compound ^{N 15 - (t-BOC)} -RA-1
From 1.7 g of pure compound RA-A-1 0.22 g.

The factor RA-A-1 / B ₀ is obtained by the same procedure as above, except that in the purification by reverse phase chromatography, only the fractions containing the pure single factor of interest are collected.

Method B 50 g of the compound of Example 2 (RA) in 200 ml of DMF (about 27 mmo
l) to the stirred solution, 3,3- (N, N-dimethylamino) -1
-Propylamine 11 ml (about 90 mmol) and PyBOP 18 g (about
35 mmol) is added at room temperature. After stirring for 15 minutes, 1 liter of ethyl acetate was added, and the precipitated solid matter (about 63 g) was collected and purified by reverse phase column chromatography.
Fractions containing pure single factor are all combined together to give 25 g of compound RA-A-1.

Example 6 Creating compound 4 (MA-A-2 / B 0) (Y is -COOCH _3, X is _{-NH- (CH 2) 3 - [} NH-
(CH _{2) 3 -]} is 2 -NH _2, R ₁ is -H, R ₂ is 9
- a methyldecyl, M is alpha-D-mannopyranosyl, Z is a compound of formula (I) when it is -H) Step ^{a: N 15 - (t-} BOC) -MA Create cyanomethyl ester dimethyl sulfoxide Example 2 in 10 ml of (DMSO),
Compound of Step ^{a (N 15 - (t-} BOC) -MA) 2.5g, TEA0.2
A solution of 5 ml and 2.5 ml of chloroacetonitrile is stirred for 4 hours at room temperature. Then 90 ml of ethyl acetate was added and the precipitated solid was collected to give the title compound N ^15-
2.8 g of (t-BOC) -MA cyanomethyl ester are obtained.

Step ^{b: N 15 - (t-} BOC) -MA-A-2 Creating the crude cyanomethyl ester compound is dissolved in DMSO30ml. The obtained solution was added with N, N, -bis- (3-
2.8 ml of aminopropyl) -1,3-propanediamine are added and the reaction mixture is stirred for 4 hours at room temperature. Thereafter, ethyl acetate 200ml was added, the solid is collected to precipitate the crude title compound ^{N 15 - (t-BOC)} -MA-A-
2 g of 3 g are obtained.

Step c: The above crude compound is the above-mentioned Example 3, Method A, Step b.
Treated with TFA as described in, and after reverse phase column chromatography, only the fractions containing pure single factor of interest were combined to give pure compound MA.
-A-2 / B ₀ 0.45 g is obtained.

Example 7: Creating a compound 5 (MA-A-3 / B 0) (Y is -COOCH _3, X is _{-NH- (CH 2) 3 -N [} -
(CH ₂₎ a ₃ -NH _{2] 2,} R ₁ is -H, R ₂ is 9-
A compound of formula (I) when it is methyldecyl, M is α-D-mannopyranosyl and Z is —H) Step a: N ′, N ″ -di (t-BOC) -tris (3-amino Propyl) amine preparation N ', N "-protected polyamines are disclosed in WO 90/113
Created as described in Issue 00.

Step b: Condensation of MA with N ', N "-di (t-BOC) -tris (3-aminopropyl) amine 18 g of the compound of Example 1 (MA) (about 10 mm) in 150 ml DMSO.
ol), protected amine 14g (about 36mmol), TEA 3ml (about 22mmo
l), and 6 ml of DPPA (about 28 mmol) are stirred for 2 hours at room temperature, then 500 ml of ethyl acetate are added. The solid that precipitates is collected (about 22 g) and used for the next step without further purification.

Step c: Removal of t-BOC-protecting group The crude product of step b was dried TF precooled at 0 ° C.
Dissolved in 150 ml A, the solution is stirred for 20 minutes at 0-5 ° C. Then 150 ml of methanol and 300 ml of diethyl ether are added. The precipitated solid was collected, washed several times with diethyl ether and then purified by reverse phase column chromatography, combining only those fractions containing pure single factor of interest to give compound MA- A-3 / B ₀ 9g is obtained.

Example 8: Compound 9 (RA-A-2) of the creation (Y is -CH ₂ OH, X is _{-NH- (CH 2) 3 - [} NH (CH
₂ ) ₃ ] _2- NH ₂ , R ₁ is -H, and R ₂ is A40926.
(C ₉ -C ₁₂ ) alkyl corresponding to the factor of the complex, M is α-D-mannopyranosyl, and Z is -H.
Compound) Step a of formula (I) when it ^{is: N 15 - (t-BOC} ) -RA in creating DMSO40ml cyanomethyl ester, Example 2, the compound of step b (N ¹⁵ -
(T-BOC) -RA) 8 g (about 4 mmol), TEA 0.75 ml (about 5.5 mm)
ol) and 8 ml of chloroacetonitrile are stirred at room temperature for 5 hours. 200 ml of ethyl acetate are then added and the solid that precipitates is collected to give 8.2 g of the crude title cyanomethyl ester.

Steps b and c: N ', N "-bis- (3-aminopropyl)
Condensation with -1,3-propanediamine and acidolysis of the t-BOC-protecting group The crude cyanomethyl ester from step a was dissolved in 80 ml DMSO and N, N'-bis- (3-aminopropyl) -1. , 3-
9 g of propanediamine are added. After stirring for 20 hours at room temperature, 320 ml of ethyl acetate are added. The solid that precipitates is collected and redissolved in 70 ml of ice cold dried TFA. The solution is stirred at 0 ° C. for 10 minutes and then 230 ml of cold diethyl ether are added. Precipitated solids are collected and water 2
Quickly redissolved in 00 ml. This solution was adjusted to pH 5.5 with 1 N NaOH and purified by reverse phase chromatography, combining all fractions containing pure single factor to give the pure title compound RA-A-2. To obtain 1.3 g of.

Example 9: Creating a compound 10 (RA-A-3) (Y is -CH ₂ OH, X is _{-NH- (CH 2) 3 -N [} (CH
₂ ) ₃ NH ₂ ] ₂ , R ₁ is —H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, and M is α-D-mannopyranosyl. , A compound of formula (I) when Z is —H) To a stirred solution of compound (RA) 9 (about 5 mmol) of Example 2 in 100 ml DMSO is added N ′, N ″ -di (t-BOC) -tris. -(3-Aminopropyl) amine (Example 7, step a) 7 g (about 18 mm
ol), 1.5 ml TEA and 3 ml DPPA are added at 10 ° C. 1
After stirring at 0 ° C for 1 hour and at room temperature for 4 hours, ethyl acetate 4
00 ml is added. The solid that precipitated (about 12 g) was redissolved in 80 ml of ice-cold dry TFA and the solution was
Stir for 0 minutes. Subsequently, a methanol / diethyl ether 1/1 mixture (about 300 ml) precooled to −10 ° C. is added. The solid that precipitates is collected and quickly recycled to 200 ml of water. This solution was adjusted to pH 4 with 1 N NaOH, purified by reverse phase chromatography, and all fractions containing pure single factor were combined to give 1.8% of pure title compound RA-A-3. g is obtained.

Example 10: creation of Compound 11 (A-A-1) (Y is -COOH, X is _{-NH- (CH 2) 3 -N (} CH 3)
₂ , R ₁ is -H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, and M is α-
D-mannopyranosyl, compound of formula (I) when Z is -H) Compound 6 (MA-A prepared as described in Example 3, Method B in 60 ml of tetrahydrofuran (THF). −
1) To 5 g (about 2.5 mmol) of stirred suspension, 10 ml of water and 1 NN
20 ml aOH is added at room temperature. After 30 minutes, the resulting solution was adjusted to pH 7 with 1N HCl, 150 ml of n-butanol was added, and the mixture was added to a small amount (about 20 ml) under reduced pressure at 40 ° C.
Is concentrated to. The solid that precipitated by the addition of about 200 ml of diethyl ether was collected (5.2 g), purified by reverse phase chromatography and all fractions containing pure single factor were combined to give the title compound AA-1. 2.1 g of is obtained.

Example 11: Compound 12 (PyA-A-1) prepared in (Y is -COO ^- and is, X is _{-NH- (CH 2) 3 -N (} CH 3)
₂ , R ₁ is —H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, and M is α
-D-mannopyranosyl, a compound of formula (I) when Z is P ⁺ (NC ₄ H ₈ ) ₃ Compound 6 (MA-A-1) to compound 11 (A-A-1)
Compound 12 (PyA-A-1) was obtained in 35% yield from compound 7 (PyMA-A-1) of Example 4 by operating under the same conditions as described in Example 10 for the preparation of Can be obtained at.

Example 12: a compound 13 Creating (A-A-3 / B 0) (Y is -COOH, X is _{-NH- (CH 2) 3 N [} (CH 2) 3 N
H ₂ ] ₂ , R ₁ is —H, R ₂ is 9-methyldecyl, M is α-D-mannopyranosyl, and Z is —.
Compound of formula (I) when H is) Compound 6 (MA-A-1) to compound 11 (A-A-1)
Compound 13 (AA-3 / B ₀ ) was converted to compound 5 of Example 7 (MA-) under the same conditions described in Example 10 for the preparation of
A-3 / B ₀ ) in a yield of 41%.

Example 13: creation of compound 14 (ABA-A-1) (Y is -CONHCH _3, X is _{-NH- (CH 2) 3 -N (} CH
₃ ) ₂ , R ₁ is -H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the AA40926 complex, M
There is alpha-D-mannopyranosyl, Z is a compound of formula (I) when it is -H) Step ^{a: N 15 - (t-} BOC) -A-A-1 · 6 B - creation of cyanomethyl ester To a stirred solution of 22 g (about 11 mmol) of compound 11 (AA-1) of Example 10 and ₃ g of NaHCO3 in 220 ml of a 1/1 water / dioxane mixture was added di-tert-butyl- in 20 ml of dry dioxane. A solution of 5 g of dicarbonate is added dropwise at room temperature over 10 minutes. After stirring for 2 hours at room temperature, 200 ml of water are added, the solution is adjusted to pH 3 with 1N HCl and extracted with 300 ml of n-butanol. The organic layer is separated and concentrated under reduced pressure at 35 ° C. to a small amount (about 45 ml). Solids precipitated by addition of diethyl ether (about 250ml) was recovered (crude ^{N 15 - (t-BOC)} -A-A-1 to about 20
g)), redissolved in 150 ml DMSO. After addition of 3 ml TEA and 20 ml chloroacetonitrile, the solution obtained is stirred for 5 hours at room temperature and then 500 ml ethyl acetate are added. Precipitating solid (approximately 18g of cyanomethyl ester)
Is high enough for use in the next step.

Step b: Reaction of the above 6 ^B -cyanomethyl ester with methylamine and removal of the t-BOC-protecting group A solution of 5 g of the above product in 75 ml of 25% (w / v) methylamine in ethanol at room temperature for 3 hours. After stirring, 300 ml of diethyl ether are added. The solid that precipitates (about 5.1 g) is collected and redissolved in 35 ml TFA. The resulting solution was stirred at 0 ° C. for 15 minutes and then 50 ml of methanol / diethyl ether 1/1 mixture was added to precipitate 4.5 g of crude product, which was subjected to reverse phase column chromatography. All fractions containing the pure single factor were combined and combined to give the title compound 14 (ABA-A-
1) 1.7 g is obtained.

Example 14: creation of compound 15 (ADA-A-1) (Y is _{-CONH- (CH 2) 3 -N (} CH 3) 2, X is -
_{NH- (CH 2) 3 -N (} CH 3) _2, R ₁ is -H,
R ₂ corresponds to the factor of A40926 complex (C ₉ −
C ₁₂ ) alkyl, M is α-D-mannopyranosyl and Z is —H, the compound of formula (I)) Antibiotics A40926 conjugate 7 g (about 4 mmol), 3, in DMF 70 ml.
2.5 ml of 3-dimethylamino-1-propylamine (about 20 mm
ol) and 5.2 g (about 10 mmol) of PyBOP at room temperature
After stirring for an hour, 400 ml of ethyl acetate is added.
The precipitated solid was collected, purified by reverse phase column chromatography, and all fractions containing pure single factor were combined to give 2.1 g of title compound 15 (ADA-A-1).
Is obtained.

Example 15: creation of compound 16 (PyRA-A-1) (Y is -CH ₂ OH, X is _{-NH- (CH 2) 3 -N (} C
H ₃ ) ₂ , R ₁ is —H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, M
Is α-D-mannopyranosyl, and Z is P ⁺ (NC ₄ H ₈ ) ₃
Compound of formula (I) when CH ₃ COO ⁻ ) Compound 7 prepared according to Example 4 in 20 ml of water
In a stirred solution of 400 mg (about 0.2 mmol) of (PyMA-A-1), n
4 ml butanol and 200 mg NaBH ₄ are added at room temperature. After stirring overnight at room temperature, the reaction mixture is adjusted to pH 4.5 with glacial acetic acid and extracted with 50 ml n-butanol. The organic layer is separated and the solvent is evaporated under reduced pressure at 45 ° C. The residual solid was purified by reverse phase chromatography and all fractions containing pure single factor were combined to give 175 mg of pure title compound 16 (PyRA-A-1).
Is obtained.

Example 16: creation of compound 25 (MA-A-4) (Y is -COOCH _3, X is , R ₁ is —H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, and M is α-D.
-Mannopyranosyl and the compound of formula (I) when Z is -H) The compound of Example 1 (M) in 60 ml of DMF / DMSO5 / 1 mixture.
A) To 5 g of a stirred solution, 0.3 ml of N-methyl-piperazine and 1.7 g of PyBOP are added at room temperature. After reacting for 1 hour, 140 ml of ethyl acetate was added, the precipitated solid was collected and purified by reverse phase column chromatography, all fractions containing pure single factor were combined to give the title compound MA. -1.9 g of A-4 is obtained.

Example 17: creation of compound 24 (RA-A-4) (Y is -CH ₂ OH, X is , R ₁ is —H, R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex, and M is α-D.
-A compound of formula (I) when it is mannopyranosyl and Z is -H) Under the above reaction conditions. The pure title compound RA-A-4 was obtained from 5 g of RA in exactly the same manner as described in Example 16 above.
2.7 g are obtained.

Reversed Phase Column Chromatography Pure samples of the above compounds are obtained by reverse phase column chromatography on silanized silica gel (0.063-0.2 mm; Merck). Crude product (eg 0.5g)
Is dissolved in a minimal amount of acetonitrile / water 1/1 mixture, then the solution is adjusted to pH 7 with 1 N NaOH and diluted with water until cloudy. Then a few drops of acetonitrile are added with vigorous stirring. As soon as a clear liquid was obtained, it was loaded onto a column of silanized silica gel (100 g) in water. Elution is 0 in 10 hours.
A linear gradient from 10% to 60% acetonitrile in 1N acetic acid was performed at a flow rate of about 250 ml / h, during which 20 ml fractions, collected by HPLC, were collected. When those fractions containing pure compounds of formula (I) are selected and complex compounds are desired where R ₂ is (C ₉ -C ₁₂ ) alkyl corresponding to the factor of the A40926 complex. All fractions containing pure factor are combined and the solvent is evaporated under reduced pressure at 40 ° C. with the addition of n-butanol to avoid foaming.

In a method of making a class of compounds of formula (I), an antibiotic A40926 complex is used and R ₂ is A 4
The single factor of the amide compound of formula (I) when corresponding to one of the meanings characterizing the individual factors of the complex (eg R ₂ = 9-methyldecyl) is determined by HPLC when desired. Only those fractions containing the desired pure factor are combined together and processed as described above.

The identity and structure of each single factor of the compounds of the invention are determined by HPLC analysis of each reaction product. Therefore, preliminary identification of the desired factor is obtained by comparison of the HLPC fingerprint of the A40926 complex with that of the crude reaction product (see, eg, "LFZerilli et al., R
apid Communications in Mass Spectrometry, Vol.C ₆ , 10
91992 ", reported HPLC pattern) (in this report, Factor B ₀ of A40926 complex is referred to as Factor B).

HPLC analysis is performed using a Varian Model 5500 liquid chromatograph equipped with a variable UV detector on a column Hibar (125x4 mm; Merck) prepacked with Li-Chrotpher RP-8 (5 μm). The chromatogram is recorded at 254 nm. Elution is 0.2 in 30 minutes
% Acetonitrile in 20% ammonium formate aqueous solution
Performed at a flow rate of 1.5 ml / min with a linear step gradient up to 60%. In general, all complex compounds of the invention have typical HPLC fingerprints that are similar to the characteristics of their respective A40926 complex precursors, so that compounds of the invention corresponding to the factors of precursor A40926 complex Individual factors can be easily identified by comparing the two HPLC patterns. The elution fraction of the reverse phase chromatogram containing the pure factor was
It can be isolated and recovered as described above. Furthermore, in order to establish the identity of the (C ₉ -C ₁₂ ) alkyl chain, a test sample of each fraction was evaporated as above to give a sample of the product, which was analyzed by F. Zerilli et al. By gas chromatography / mass spectrometry (GC / MS) according to the method described in the above-mentioned document.

Table V shows the retention times (t _R ) of pure factors of each invention compound of formula (I) when R ₂ is 9-methyldecyl (ie, corresponding to factor B ₀ of the A40926 complex). Which is referred to in the reverse phase purification method.

The table also shows the factors for the A40926 complex precursor.
Also shown is B ₀ and the t _{R of} the corresponding starting material (MA) ester recorded under the same conditions as above.

¹ H- and ³¹ P-NMR spectrum The ¹ H-NMR spectrum at 500 MHz was measured by the internal standard (δ
= 0.00 ppm) using tetramethylsilane (TMS) in DMSO-D _{6 at} a temperature range of 20 to 30 ° C, BrukerAM
Recorded on a 500 spectrophotometer. Table VI shows the most significant chemical shifts (δ ppm) of some representative compounds.

^{The 31} P-NMR spectrum shows DMSO-D at 161.98 MHz (compound 12), or 202.46 MHZ (compounds 7 and 16).
Recorded in ₆ solution with 85% H ₃ PO ₄ as internal standard.

Compound 7 (PyMA-A-1) ( ³¹ P): 1 at δ24.12 ppm
Signal compound 12 (PyA-A-1) ( ³¹ P): 1 at δ23.50 ppm Signal compound 16 (PyRA-A-1) ( ³¹ P): 1 at δ24.11 ppm
Signals The main features or aspects of the present invention are as follows.

1. An antibiotic A40926 derivative of formula (I) or a pharmaceutically acceptable addition salt thereof.

  In the above formula, R₁Is hydrogen or a protecting group of amino functional group
Represents; R₂Is (C₉-C₁₂) Represents alkyl; M is hydrogen,
α-D-mannopyranosyl or 6-O-acetyl-
represents α-D-mannopyranosyl; Y is carboxy,
(C₁-C_Four) Alkoxycarbonyl, aminocarbonyl,
(C₁-C_Four) Alkylaminocarbonyl, di (C₁-C_Four)
Rualkylaminocarbonyl [These alkyl moieties are
Droxy, amino, (C₁-C_Four) Alkylamino and di
(C₁-C_Four) One substituent selected from alkylamino
You may have. ] Or hydroxymethyl
X; is hydroxy or a formula -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃Is hydrogen or (C₁-C_Four) Represent alkyl
Shi; alk₁, Alk₂And alk₃Are each independently a carbon atom 2
Represents ~ 10 linear or branched alkylenes; p and q
Is independently an integer representing 0 or 1; R_FourAnd R
_FiveAre each independently hydrogen, (C₁-C_Four) Represents alkyl
Or R₃And R_FourTogether, under the condition that p is 1,
Linked to two nitrogen atoms (C₂-C_Four) Alkylene part
Represents minutes or R_FourAnd R_FiveTogether, both p and q
Linked to two nitrogen atoms under the condition that one is 1 (C₂
-C_Four) Represents an alkylene moiety; W is hydrogen, (C₁-C_Four)
Rukiru, Amino, (C₁-C_Four) Alkylamino, di (C₁−
C_Four) Alkylamino, 1 or 2 amino- (C₂−
C_Four) Alkyl moieties or 1 or 2 (C₁-C_Four)
Rukiruamino- (C₂-C_Four) Alkyl moiety or 1
Is two di (C₁-C_Four) Alkylamino- (C₂-C_Four) Al
Represents or is substituted by an amino group
Is a partial W if p and q are both zero
-NR₃−alk₁-With, piperazino or
May represent 4-methylpiperazino] In the case where X represents hydroxy,
In, Y represents hydroxymethyl, Z is hydrogen or
Is based [In the formula, A Represents an anion of an inorganic or organic acid
Or carboxylic acid functionality left on the antibiotic
If present in the part, it also includes the carboxylic acid
May represent an internal anion derived from a functional group] Represents

2.R₁Represents a protecting group for hydrogen or an amino functional group; R₂
But (C₉-C₁₂) Represents alkyl; M is hydrogen, α-D-
Mannopyranosyl or 6-O-acetyl-α-D-
Represents mannopyranosyl; Y is carboxy, (C₁-C_Four)
Alkoxycarbonyl, aminocarbonyl, (C₁-C_Four)
Alkylaminocarbonyl, di (C₁-C_Four) Alkylami
Nocarbonyl, the alkyl part of which is
Roxy, amino, (C₁-C_Four) Alkylamino and di
(C₁-C_Four) Also one substituent selected from alkylamino
Or X may be hydroxymethyl.
Shi or formula -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_FourAnd R_FiveRepresents hydrogen; alk₁, Alk₂Oh
And alk₃Are each independently linear with 2 to 4 carbon atoms.
Represents a branched alkylene; p and q are independently 0 or
Is an integer representing 1; W is hydrogen, (C₁-C_Four) Al
Kill, amino, (C₁-C_Four) Alkylamino, di (C₁−
C_Four) Alkylamino, 1 or 2 amino- (C₂−
C_Four) Alkyl moieties or 1 or 2 (C₁-C_Four)
Rukiruamino- (C₂-C_Four) Alkyl moiety or 1
Is two di (C₁-C_Four) Alkylamino- (C₂-C_Four) Al
Represents an amino substituted by a kill moiety, or
If p and q are both zero, then W is a partial −NR
₃−alk₁With-, piperazino or 4-
May represent methylpiperazino] In the case where X represents hydroxy,
In which Y represents hydroxymethyl; Z is hydrogen or
Basis [In the formula, A Is an anion of an inorganic or organic acid, or
Is a carboxylic acid functional group on the remaining part of the antibiotic.
If present, it is also derived from the carboxylic acid functionality.
May represent internal anions that are induced] The antibiotic A40926 derivatives according to item 1, which represents

3. R ₂ represents (C ₁₀ -C ₁₁ ) alkyl, M represents α-D-mannopyranosyl, and R ₁ , X, Y and Z are the antibiotic A40926 derivatives according to item 1 above.

4. R ₁ represents hydrogen or a protecting group for an amino functional group, preferably hydrogen; R ₂ represents 7-methyloctyl, n-nonyl, 8-methylnonyl, n-decyl, 9-methyldecyl, n-undecyl or represents n-dodecyl; M is
Represents hydrogen or α-D-mannopyranosyl; Y is
Carboxy, (C ₁ -C ₄ ) alkoxycarbonyl, aminocarbonyl, (C ₁ -C ₄ ) alkylaminocarbonyl and di (C ₁ -C ₄ ) alkylaminocarbonyl are represented, and the alkyl moiety thereof is hydroxy or amino. , May have one substituent selected from (C ₁ -C ₄ ) alkylamino and di (C ₁ -C ₄ ) alkylamino or hydroxymethyl; X is —NR ₃ —alk ₁ — (NH -Alk ₂ ) p- (NH-alk ₃ ) q-W [In the formula, R ₃ represents hydrogen; alk ₁ , alk ₂ and alk ₃ are
Each independently represents a linear alkylene having 2 to 4 carbon atoms; p and q each independently represent 0 or 1, and W is amino, (C ₁ -C ₄ ) alkylamino, di ( C ₁
-C ₄ ) alkylamino, 1 or 2 amino- (C ₂
-C ₄₎ an amino substituted by alkyl moiety,
Alternatively, when p and q are both zero, W is, together with the moiety -NR ₃ -alk _1- represents piperazino or 4-methylpiperazino], and Z is The antibiotic A409 according to above 1, which represents hydrogen.
26 derivatives.

5. R ₁ represents hydrogen; R ₂ is 7-methyloctyl, n-
Represents nonyl, 8-methylnonyl, n-decyl, 9-methyldecyl, n-undecyl or n-dodecyl; M
Represents α-D-mannopyranosyl; Y represents carboxy, methoxycarbonyl, aminocarbonyl, methylaminocarbonyl, dimethylaminocarbonyl, (dimethylamino) ethylaminocarbonyl or hydroxymethyl; X represents —NH— (CH _{2) 3 -N (CH 3)} 2, -NH (CH 2) 3 - [NH- (CH 2) 3] 2 -NH 2, -NH- (CH 2) 3 -N [(CH 2) 3 NH ₂ ] ₂ and The antibiotic A40926 antibiotics according to item 1 above, which is an amino residue selected from: Z represents hydrogen.

6. R ₂ represents n-decyl, 8-methylnonyl, 9-methyldecyl or n-undecyl; R ₁ , M, Y, X and Z
Is the antibiotic A40926 derivative stream according to item 5 above, as described in item 5 above.

7. R ₂ represents 9-methyldecyl; R ₁ , M, Y, X and Z
Is the antibiotic A40926 derivative according to item 5 above, as described in item 5 above.

8. Y is methoxycarbonyl or hydroxymethyl; X is —NH— (CH ₂ ) ₃ —N (CH ₃ ) ₂ ; R ₁ ,
Antibiotic A40926 derivatives according to item 5 above, wherein R ₂ , M and Z are as described in item 5 above.

9. Antibiotics A40926 derivatives of formula (I) and their pharmaceutically acceptable addition salts.

In the above formula, R ₁ represents hydrogen or a protecting group for an amino functional group; R ₂ represents (C ₁₀ -C ₁₁ ) alkyl; M represents hydrogen, α-D-mannopyranosyl or 6-O- Represents acetyl-α-D-mannopyranosyl; Y is (C ₁ -C ₄ );
Represents an alkoxycarbonyl or hydroxymethyl; X represents hydroxy or the formula -NR ₃ -alk _1- (NR ₄ -alk ₂ ) p- (NR ₅ -alk ₃ ) q-W [wherein R ₃ is hydrogen or Represents (C ₁ -C ₄ ) alkyl; alk ₁ , alk ₂ and alk ₃ are each independently a carbon atom 2;
Represents ~ 10 linear or branched alkylenes; p and q
Is independently an integer representing 0 or 1; R ₄ and R
₅ each independently represents a hydrogen atom, (C ₁ -C ₄ ) alkyl, or R ₃ and R ₄ are taken together and are linked to two nitrogen atoms under the condition that p is 1. Represents a (C ₂ -C ₄ ) alkylene moiety or R ₄ and R ₅ are taken together and are linked to two nitrogen atoms provided that both p and q are 1 (C ₂ −
C ₄₎ alkylene moiety; W is hydrogen, (C ₁ -C ₄₎ alkyl, amino, (C ₁ -C ₄₎ alkylamino, di (C ₁ -
C ₄₎ alkylamino, 1 or 2 amino - (C ₂ -
C ₄ ) alkyl moiety or 1 or 2 (C ₁ -C ₄ ) alkylamino- (C ₂ -C ₄ ) alkyl moieties or 1 or 2 di (C ₁ -C ₄ ) alkylamino- (C ₂ -C ₄₎ amino acid residues of the alkyl moiety represents an amino substituted by] are provided that if X represents hydroxy, Y represents hydroxymethyl.

10.R ₁ represents hydrogen or a protecting group for an amino functional group;
R ₂ represents (C ₁₀ -C ₁₁ ) alkyl; M is hydrogen, α-D
-Mannopyranosyl or 6-O-acetyl-α-D
- represents mannopyranosyl; Y is, (C ₁ -C ₄₎ alkoxycarbonyl or hydroxymethyl; X is hydroxy or of the formula _{-NR 3 -alk 1 - (NR 4} -alk 2) p- (NR 5 -alk ₃ ) q-W [wherein R ₃ , R ₄ and R ₅ represent hydrogen; alk ₁ , alk ₂ and alk ₃ are each independently a linear or branched alkylene having 2 to 4 carbon atoms. Represents an integer; p and q are each independently an integer representing 0 or 1; W represents hydrogen, (C ₁ -C ₄ ) alkyl, amino, (C ₁ -C ₄ ) alkylamino, di (C ₁ −
C ₄ ) alkylamino, or 1 or 2 amino-
(C ₂ -C ₄ ) alkyl moiety or 1 or 2 di (C ₁
-C ₄ ) alkylamino- (C ₂ -C ₄ ) represents an amino substituted by an alkyl moiety], provided that when X represents hydroxy, Y represents hydroxymethyl. The antibacterial substance according to item 9.

11. The antibacterial substance as described in 9 above, wherein R ₂ represents 9-methyldecyl, M represents α-D-mannopyranosyl, and R ₁ , X and Y are as described in 9 above. .

12.R ₂ represents (C ₁₀ -C ₁₁ ) alkyl, preferably 9-methyldecyl; Y represents (C ₁ -C ₄ ) alkoxycarbonyl, preferably methoxymethyl or hydroxymethyl, and X represents _{-NH- (CH 2) 3 -N (} CH 3) 2, -NH- (CH 2) 3 - [NH- (CH 2) 3] 2 -NH 2 and -NH- (CH _{2) 3} -N [ 10. The antibacterial substance as described in 9 above, which is selected from (CH ₂ ) ₃ NH ₂ ] ₂ .

13. Formula (I)   In the above formula, R₁Is hydrogen or a protecting group of amino functional group
Represents R₂Is (C₉-C₁₂) Represents alkyl; M is water
Elemental, α-D-mannopyranosyl or 6-O-acetyl
Represents -alpha-D-mannopyranosyl; Y is carboxy
Si, (C₁-C_Four) Alkoxycarbonyl, aminocarboni
Le, (C₁-C_Four) Alkylaminocarbonyl, di (C₁−
C_Four) Alkylaminocarbonyl [These alkyl moieties
Is hydroxy, amino, (C₁-C_Four) Alkylamino
And J (C₁-C_Four) One unit selected from alkylamino
It may have a substituent. ] Or hydroxymethyl
Represents X is hydroxy or a formula -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃Is hydrogen or (C₁-C_Four) Represent alkyl
Shi; alk₁, Alk₂And alk₃Are each independently a carbon atom 2
Represents ~ 10 linear or branched alkylenes; p and q
Is independently an integer representing 0 or 1; R_FourAnd R
_FiveAre each independently hydrogen, (C₁-C_Four) Represents alkyl
Or R₃And R_FourTogether, under the condition that p is 1,
Linked to two nitrogen atoms (C₂-C_Four) Alkylene part
Represents minutes or R_FourAnd R_FiveTogether, both p and q
Linked to two nitrogen atoms under the condition that one is 1 (C₂
-C_Four) Represents an alkylene moiety; W is hydrogen, (C₁-C_Four)
Rukiru, Amino, (C₁-C_Four) Alkylamino, di (C₁−
C_Four) Alkylamino, 1 or 2 amino- (C₂−
C_Four) Alkyl moieties or 1 or 2 (C₁-C_Four)
Rukiruamino- (C₂-C_Four) Alkyl moiety or 1
Is two di (C₁-C_Four) Alkylamino- (C₂-C_Four) Al
Represents or is substituted by an amino group
Is a partial W if p and q are both zero
-NR₃−alk₁-With, piperazino or
May represent 4-methylpiperazino] In the case where X represents hydroxy,
In, Y represents hydroxymethyl, Z is hydrogen or
Is based [In the formula, A Represents an anion of an inorganic or organic acid
Or carboxylic acid functionality left on the antibiotic
If present in the part, it also includes the carboxylic acid
May represent an internal anion derived from a functional group] A method for producing an antibiotic A40926 derivative that represents (A) R₁, R₂, M, Y and Z are as defined above
Yes, and X is an amino residue -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_Four, R_Five, Alk₁, Alk₂, Alk₃, P, q and
W is as indicated at the beginning of this claim]
The compounds of formula (I) in some cases may, if desired, be of formula
(II) [In the above formula, R '₁, R '₂And M'is R₁, R₂And same as M
And Y'is (C₁-C_Four) Is an alkoxycarbonyl,
And X ′ is hydroxy], the compound of formula (III) [In the formula, R₃, R_Four, R_Five, Alk₁, Alk₂, Alk₃, P, q and
W is as described above] and the condensing agent
Present or C⁶³Of "active ester" of carboxylic acid
Through formation, undergoing amidation reaction, and i) optionally Y is (C₁-C_Four) Alkoxycarbonyl
And then R₁Is N¹⁵-A suitable protecting group for the amino function
And all other symbols are as defined above
The resulting derivative of formula (I)
Reduction with a metal and optionally Y is a hydroxy
Simethyl and R₁Gives the corresponding compound in which is hydrogen
For N¹⁵Removing the protecting group; ii) Optionally, Y is (C₁-C_Four) Alkoxycarbonyl
Is hydroxymethyl and R₁Is N¹⁵-Amino functional group
A suitable protecting group for R and₂, M and Z are the same as above.
Is as defined, and X is an amino residue-NR₃−alk₁-NHR_Four
[In the formula, R₃, R_FourAre each independently hydrogen or (C₁−
C_Four) Represents alkyl, and alk₁Is as above
Or] or   -NR₃−alk₁-NR_Four−alk₂-NHR_Five [In the formula, R₃, R_Four, R_FiveAre each independently hydrogen or (C₁
-C_Four) Represents alkyl, and alk₁And alk₂Is above
Is as follows] The resulting derivative of formula (I) when is also of formula (IV)
(IV a), respectively [In the formula, R_Five, Alk₂, Alk₃And W are as described above
, Q is 0 or 1, and r is halo, methan
Represents a sulfonyl or tosyl] amine reactant
Alkylate in the presence of an acid acceptor in an inert solvent,
And, optionally, N¹⁵Removing the protecting group; iii) optionally, Y is (C₁-C_Four) With alkoxycarbonyl
Yes, and other symbols are as defined above.
The resulting derivative of formula (I) in which Y is carboxy
To obtain the corresponding compound,
Treated with a solution; iiii) optionally M is α-D-mannopyranosyl or
Is 6-O-acetyl-α-D-mannopyranosyl.
And other symbols are as defined above
The resulting derivative of formula (I) in which the corresponding M is hydrogen
Acid hydrolysis to give the compound (B) R₁, R₂, X and M are as defined above.
And Y is hydroxymethyl and Z is hydrogen.
If a compound of formula (I) of₁But N¹⁵−
A suitable protecting group for the amino functional group, R '₂But R₂Same as
And Y'is (C₁-C_Four) Is an alkoxycarbonyl,
And a compound of formula (II) where X'is hydroxy is hydrogen
Reduction with alkali metal borohydride, and optionally,
R₁To obtain the corresponding compound in which is hydrogen¹⁵− Protection
Remove the guardian, and i) optionally Y is hydroxymethyl and R₁Is hydro
Xy and all other symbols are as defined above.
The resulting compound of formula (I) in certain cases is: Formula (III) -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_Four, R_Five, Alk₁, Alk₂, Alk₃, P, q and
W is as described above].
C in the presence of a mixture or in an inert organic solvent⁶³Carbo
The amidation reaction is carried out through the formation of the "active ester" of the acid.
Performance; (C) R₁, R₂, M and Z are as defined above.
Y and the partial COX are the same group (C₁-C_Four) Alkyla
Minocarbonyl, di (C₁-C_Four) Alkylaminocarboni
And the alkyl moiety is amino, (C₁−
C_Four) Alkylamino and di (C₁-C_Four) Alkylamino
Induction of formula (I) when it may have a substituent selected from
R'if a conductor is desired₁, R '₂And M'is R₁, R₂
And M are the same, Y'is carboxy, and
Compounds of formula (II) wherein X'is hydroxy are
As in (a), the symbol R₃, R_Four, R_Five, Alk₁, Alk₂, P,
q and W are the carboxamide residues Y and Y as defined above.
And a selection of formula (III) above with the same appropriate meaning as COX
Carried out an amidation reaction with excess amine prepared; (D) R₁, R₂, M and Z are as defined above.
, Y and partial COX have different carboxamide residues
Representing, the meaning of Y is aminocarbonyl, (C₁-C_Four)
Rukiruaminocarbonyl, di (C₁-C_Four) Alkylamino
Selected from carbonyl, the alkyl moiety is hydro
Xy, amino, (C₁-C_Four) Alkylamino and di (C₁
-C_Four) May have a substituent selected from alkylamino
And the meaning of X is the formula -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_Four, R_Five, Alk₁, Alk₂, Alk₃, P, q and
W has the same meaning as in the beginning of this claim] Derivatives of formula (I) when amino residues of
in case of: i) R₁, R₂, M and Z are as described above, and X is
Amino residue -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_Four, R_Five, Alk₁, Alk₂, Alk₃, P, q and
W is as described above] And a derivative of formula (I) when Y is carboxy
Forms a carboxamide residue Y as defined above
For the amidation with a suitable amine in the presence of a condensing agent
Do the reaction, or ii) R₁Is N¹⁵-Represents a protecting group for an amino functional group, R₂, M
And Z are as described above, and X is an amino residue. -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [In the formula, R₃, R_Four, R_Five, Alk₁, Alk₂, Alk₃, P, q and
W is as described above] and Y is carboxy
In some cases the derivative of formula (I) may be at position 6^BCorrespondence in
Carboxa as defined above.
Suitable with its active ester to form the mid residue Y
A suitable amine, and optionally R₁Is hydrogen
N to obtain the corresponding compound¹⁵-Eliminating the protecting group
A method characterized by:

14. Steps (a), (b) i), (c) and (d) i)
In the amidation operation in (C ₁
-C ₄ ) carried out in the presence of a condensing agent selected from alkyl, phenyl, heterocyclic azidophosphate and benzotriazolyloxy-tris- (pyrrolidino) phosphonium hexafluorophosphate at a temperature of 0 ° C to 20 ° C. The method of paragraph 13 above, further characterized in that

15. Steps (a), (b) i), (c) and (d) ii)
Amidation operation of the carboxylic acid starting material of formula (II),
Carried out by converting to the corresponding active ester, preferably protected at the N ¹⁵ -amino function,
It is a further feature that the active ester is reacted with a molar excess of the amine of formula (III) in the presence of an organic polar solvent at a temperature of 5 ° C to 60 ° C, preferably 10 ° C to 30 ° C. The method according to paragraph 13 above.

16. The active ester is cyanomethyl ester, the molar amount of which is 1: 5-1: 30 with respect to the amine,
The method of paragraph 15 above, which is further characterized.

17. The cyanoester is protected with the carboxylic acid starting material of formula (II), preferably the N ¹⁵ -amino functional group, to give 20-3
It is prepared by reacting with a 0-fold molar excess of chloroacetonitrile in the presence of an inert organic solvent and a base that does not interfere with the progress of the reaction at a temperature of 5 ° C to 60 ° C, preferably 10 ° C to 30 ° C. The method according to paragraph 16 above, further characterized by:

18. In steps (a) i) and (b), the molar ratio of alkali metal borohydride to the compound of formula (II) is from 50 to 30.
A process according to paragraph 13 further characterized in that it is comprised between 0 and the reaction is carried out at a temperature between 0 ° C and 40 ° C, preferably at room temperature.

19. The inert organic solvent used is dimethylformamide, dimethoxyethane, hexamethylphosphoramide,
15. The method according to paragraph 14, further characterized by being selected from dimethyl sulfoxide and mixtures thereof.

20. The organic polar solvent is selected from lower alkanols, dimethylformamide, hexamethylphosphoramide, 1,3-dimethyl-3,4,5,6-tetrahydro-2 (1H) -pyrimidone, dimethyl sulfoxide and dimethoxyethane. 16. The method of paragraph 15 above, further characterized in that:

21. The alkali metal borohydride is selected from sodium borohydride, potassium borohydride and sodium cyanoborohydride, and its reaction solvent is a mixed solution of water and a water-soluble or partially soluble lower alkanol. The method according to Item 18, wherein diethyl ether is optionally added as a defoaming agent.

22.N ¹⁵ - protecting group, t-butoxy carbonyl or carbobenzyloxy group, the protecting group, to be finally removed any amidation operation, according to the above item 13 which further characterized Method.

23. One or more amino groups of the amine of formula (III) not involved in the formation of an amide bond are protected before the amine is involved in the amidation reaction and deprotected after the reaction is complete. Which is further characterized in that
The method described in paragraph 13.

24.a) A compound of formula (I) wherein Y is hydroxymethyl, X is hydroxy and R ₁ , R ₂ and M are as defined in claim 9: Formula (II) if desired [In the above formula, X 'is hydroxy, Y' is, (C ₁ -C ₄₎
Alkoxycarbonyl and R ′ ₁ is a suitable protecting group for the amino functional group] at a temperature of 0-40 ° C. in a water or aqueous alcohol medium, preferably sodium borohydride, borohydride. Reducing with an alkali metal borohydride selected from potassium and sodium cyanoborohydride, and optionally removing protecting groups, b) X is a moiety -NR ₃ -alk _1- (NR ₄ -alk ₂ ) p- (NR
₅₋ alk ₃ ) q−W, and represents Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , alk ₁ , alk
_{If 2} , alk ₃ , p, q and M are compounds of formula (I) when as described in paragraph 9 above, then i) X ′ is hydroxy and Y ′ is , (C ₁ -C ₄ ) alkoxycarbonyl, R ′ ₁ is a suitable protecting group for the amino functionality, a carboxylic acid compound of formula (II), or R ₂ and M are as described above, A carboxylic acid compound of formula (I) wherein Y is hydroxymethyl, X is hydroxy and R ₁ is a suitable protecting group for the amino functional group;
Formula (III) An excess of a suitable amine of the formula: wherein R ₃ , R ₄ , R ₅ , alk ₁ , alk ₂ , alk ₃ , p, q and W have the same meanings above, in an inert organic solvent. At a temperature of 0 ° C to 20 ° C in the presence of a condensing agent selected from (C ₁ -C ₄ ) alkyl, phenyl or heterocyclic azidophosphate.
And optionally removing the protecting group of the amino functional group, or ii) converting the carboxylic acid compound of formula (II) or (I) as defined above into its corresponding active ester, which active ester And the amine (III) in the presence of an organic polar solvent at a temperature of 5 to 60 ° C., preferably 10 to 30
The method according to item 9 above, wherein the reaction is carried out at 0 ° C. and, optionally, the protecting group of the amino functional group is removed.

25. The substance according to any one of items 1 to 12 above for use as a pharmaceutical.

26. 1 to 1 above regarding the manufacture of pharmaceuticals for protection against bacterial infection
The substance according to any one of item 2.

27. A pharmaceutical composition containing the compound according to any one of items 1 to 12 above as an active ingredient.

─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Pantsone, Giambatteista Milan of Italy, 20010 Cornaledo Biavanzago 23 (72) Inventor Maratsui, Aletsandra Maria Varese, Italy 21047 Salonno, Biapiave 73 (56) References JP-A-1-50900 (JP, A) (58) Fields investigated (Int.Cl. ⁷ , DB name) CA (STN) REGISTRY (STN) BIOSIS / WPI (DIALOG)

Claims (5)

(57) [Claims] 1. An antibiotic A40926 derivative of the following formula (I):
Or its pharmaceutically acceptable addition salts: In the above formula, R₁Is a protective group for hydrogen or an amino functional group.
Representation; R₂Is (C₉-C₁₂) Represents alkyl; M is hydrogen, α
-D-mannopyranosyl or 6-O-acetyl-α
Represents -D-mannopyranosyl; Y is carboxyl,
(C₁-C_Four) Alkoxycarbonyl or hydroxy
Represents chill; X is an expression -NR₃−alk₁-(NR_Four−alk₂) P- (NR_Five−alk₃) Q-W [Where R₃Is hydrogen or (C₁-C_Four) Represent alkyl
Shi; alk₁, Alk₂And alk₃Are each independently a carbon atom 2
Represents ~ 10 linear or branched alkylenes; p and q
Is independently an integer representing 0 or 1; R_FourAnd R
_FiveAre each independently hydrogen, (C₁-C_Four) Represents alkyl
Or R₃And R_FourTogether, under the condition that p is 1,
Linked to two nitrogen atoms (C₂-C_Four) Alkylene part
Represents minutes or R_FourAnd R_FiveTogether, both p and q
Linked to two nitrogen atoms under the condition that one is 1 (C₂
-C_Four) Represents an alkylene moiety; W is hydrogen, (C₁-C_Four)
Rukiru, Amino, (C₁-C_Four) Alkylamino, di (C₁−
C_Four) Alkylamino, 1 or 2 amino- (C₂−
C_Four) Alkyl moieties or 1 or 2 (C₁-C_Four)
Rukiruamino- (C₂-C_Four) Alkyl moiety or 1
Is two di (C₁-C_Four) Alkylamino- (C₂-C_Four) Al
Represents or is substituted by an amino group
Is a partial W if p and q are both zero
-NR₃−alk₁-With, piperazino or
May represent 4-methylpiperazino] Represents an amino residue of, Z is hydrogen or a group [Where A Represents an anion of an inorganic or organic acid.
Or the carboxylic acid functional group is
If present in the part, it also includes the carboxylic acid
May represent an internal anion derived from a functional group] Represents 2. The antibiotic A40926 derivative or the pharmaceutically acceptable addition salt thereof according to claim 1, wherein Y is carboxyl. 3. R ₁ represents hydrogen; R ₂ represents 7-methyloctyl, n-nonyl, 8-methylnonyl, n-decyl, 9
- methyldecyl, n- represents undecyl or n- dodecyl; M is represents alpha-D-mannopyranosyl; Y is a carboxyl; X _{is, -NH- (CH 2) 3 -N} (CH 3) 2, _{-NH (CH 2) 3 - [} NH- (CH 2) 3] 2 -NH 2, -NH- (CH 2) 3 -N [(CH 2) 3 NH 2] 2 and The antibiotic A40926 derivative or a pharmaceutically acceptable addition salt thereof according to claim 1, wherein Z is hydrogen. 4. The antibiotic A40926 derivative or a pharmaceutically acceptable addition salt thereof according to any one of claims 1 to 3, wherein X is —NH— (CH ₂ ) ₃ —N (CH ₃ ) ₂ . 5. A pharmaceutical preparation for protecting a bacterial infection, which comprises the antibiotic A40926 derivative or the pharmaceutically acceptable addition salt thereof according to any one of claims 1 to 4 as an active ingredient.